Improved light transmissive sample holder and analysis, in particular for hemoglobin

Abstract

Wherein the invention relates to a device and a method for improving the optical analysis of a sample thin layer of a sample sandwiched between two plates.

Description

[0001] This application is a divisional application filed with the Chinese application number 2018800893972, filed on December 14, 2018, and entitled "Improved Light Transmission Sample Holder and Analysis, Particularly for Hemoglobin". This application claims the benefit of U.S. Provisional Patent Application No. 62 / 598,899 filed on December 14, 2017. For all purposes, the contents of which are relied upon and incorporated herein by reference in their entirety. Technical Field

[0002] The present invention relates to an apparatus and method for improving light transmission analysis of a thin layer of a sample sandwiched between two plates. Technical Background

[0003] The absorption of light by a thin layer of a sample is one of the methods used to measure the properties of biological and chemical samples. One method of measuring the absorption of light is to measure the intensity of the incident and transmitted light directly into and out of the sample.

[0004] However, for a variety of reasons, it can be difficult to measure these light intensities directly in many practical situations. One reason is that thin layer samples often require a sample holder for measurement, and the transmitted light being measured is the light that passes through both the sample and the sample holder. Therefore, a method is needed that can separate the light absorbed by the sample holder from the light absorbed by the sample holder.

[0005] Another reason is that both the incident light and the transmitted light are on the other side of the sample, so it is difficult to detect both lights at the same time using a single detector. Therefore, it becomes a necessity to use a single photodetector for absorption measurement.

[0006] In existing methods for light transmission measurement of thin samples, a sample holder including two plates has been used to sandwich the sample in a thin layer between the two plates, and light transmission through an air bubble inside the sample thin layer (which may occur under certain conditions) is used as a reference signal to separate the light absorption of the sample holder from that of the sample. This method also allows light absorption measurement using a single photodetector. In this method, it is assumed that (i) the light transmission through the bubble region is the same as the light transmission through the zero-thickness sample, and (ii) the light absorption of the sample holder is the same in the bubble region (where the reference signal is taken) and the sample region (where a single sample is measured). However, in reality, both assumptions may be wrong. Bubbles may be generated far away from the sample signal location, so there is a significant difference in the sample holder absorbance between the two locations. The bubble may be so small that the light will be significantly scattered and the reference signal is significantly different from the sample with zero thickness. In addition, air bubble generation has randomness in the generation (generation or may not be generated) and the location of generation (e.g., random location).

[0007] Therefore, an object of the present invention is to provide an apparatus and method for generating reference light, simplifying light transmission measurement and simplifying sample handling. The present invention can overcome or reduce the disadvantages of existing devices or systems. Summary of the invention

[0008] The present invention relates to an apparatus and method for improving the optical analysis of a thin layer of a sample sandwiched between two plates, and more particularly to an apparatus and method for generating a reference signal for improving the optical analysis and for analyzing hemoglobin.

[0009] The properties of a sample (e.g., biological or chemical properties) can be determined by the optical density (i.e., OD) of the sample, the light transmitted through a thin sample layer versus the incident light (i.e., Beer-Lambert law). However, thin layer samples usually require a sample holder for measurement, and the light being measured also passes through the sample holder. The light transmission signal and light absorption (e.g., optical density) of the sample need to be separated from the total transmitted light, which includes light transmission through the sample and through the sample holder.

[0010] It is an object of the present invention to provide devices and methods of certain embodiments of sample holders and methods of using the same to improve light transmission analysis.

[0011] The details are as follows:

[0012] 1. A device comprising:

[0013] The first board, the second board, the light guide spacer (LGS), the sampling area and the reference area, where:

[0014] (i) the first plate and the second plate are configured to sandwich a sample in a thin layer between the plates for light transmission analysis, and each plate has a sample contact area on its inner surface for contact with the sample;

[0015] (ii) The light guide spacer (LGS) has a cylindrical shape and is sandwiched between the two plates, with both ends of the column directly contacting each plate to form the LGS-plate contact area, thereby forming a structure that allows light to be transmitted from the first plate through the LGS to the second plate without passing through the sample,

[0016] (iii) a sampling region is a region where light can sequentially pass through the first plate, the sample, and the second plate, wherein the sampling region does not have an LGS; and

[0017] (iv) the reference area is the area where light passes through the first plate, the light guide spacer, and the second plate in sequence without passing through the sample;

[0018] Among them, the LGS contact surface and the cross-sectional scale of LGS are larger than the wavelength of light.

[0019] wherein the light guide spacer is surrounded by the sample or is adjacent to the sample; and

[0020] Here, the thickness of the sample in the sampling area is 500 μm or less.

[0021] 2. A device for analyzing hemoglobin in a sample, comprising:

[0022] A first board, a second board and a light guide spacer, wherein:

[0023] (i) the first panel and the second panel are movable relative to each other to form different configurations, including an open configuration and a closed configuration;

[0024] (ii) each plate comprises an inner surface having a sample contact area for contacting a sample containing or potentially containing hemoglobin; and

[0025] (iii) the light-guiding gasket has a columnar shape, wherein the top surface and the bottom surface of the light-guiding gasket are substantially flat, and the bottom surface is fixed to the inner surface of one of the plates, wherein the bottom surface and the average cross-section of each gasket are respectively larger than the wavelength of light of the analyzed sample, and wherein the light-guiding gasket is within the sample contact area.

[0026] Among them, an open configuration is a configuration in which: the two plates are separated, the spacing between the plates is not limited by the light-guiding spacer, and the sample is deposited on one or both plates.

[0027] wherein the closed configuration is the configuration of the sample after deposition in the open configuration; in the closed state: at least a portion of the sample is compressed by the two plates into a highly uniform layer, wherein the uniform thickness of the layer is limited by the plates and the light guide spacers in the sample area of ​​the plates; and

[0028] wherein, in a closed configuration: (a) at least one gasket in the sample contact area has its top surface in direct contact with one of the plates, and at least one gasket and areas on the plates above and below it define a reference area, wherein the reference area is transparent to light within a specific wavelength range, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate are not occupied by a light-guiding gasket, thereby defining a sampling area that is transparent to light in the same specific wavelength range.

[0029] 3. A device for analyzing hemoglobin in an analyte in a sample, comprising:

[0030] A first board, a second board and a light guide spacer, wherein:

[0031] (i) the first plate and the second plate are configured to receive a sample that contains or may contain an analyte, wherein at least a portion of the sample is between and in contact with the two plates; and

[0032] (ii) the light guide spacer has a columnar shape and a predetermined basic height,

[0033] wherein the top and bottom surfaces of the light-guiding spacers are substantially flat, and the top and bottom surfaces of at least one spacer are in direct contact with the board,

[0034] wherein the top and bottom surfaces of each spacer and the average cross-section thereof are larger than the wavelength of light used to analyze the sample, and

[0035] wherein: (a) at least one spacer and areas of the plates above and below it define a reference area that is transparent to light within a specific wavelength range and through which light can pass through the plates and spacers, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate is not occupied by a light-guiding spacer, thereby defining a sampling area that is transparent to light within the same wavelength range.

[0036] 4. A device for analyzing an analyte in a sample, comprising:

[0037] A first board, a second board and a light guide spacer, wherein:

[0038] (i) the first panel and the second panel are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0039] (ii) each plate comprises an inner surface having a sample contact area for contacting a sample that contains or may contain an analyte; and

[0040] (iii) the light-guiding spacers have a cylindrical shape and have a predetermined substantially uniform height, wherein the top and bottom surfaces of the light-guiding spacers are substantially flat and the bottom surface is secured to the inner surface of the one or more plates, wherein the top and bottom surfaces and the average cross-sectional dimension of each spacer are respectively larger than the wavelength of light for analyzing the sample, wherein at least one of the light-guiding spacers is within the sample contact region;

[0041] Among them, an open configuration is a configuration in which: the two plates are separated, the spacing between the plates is not limited by the light-guiding spacer, and the sample is deposited on one or both plates.

[0042] wherein the closed configuration is the configuration formed after the sample is deposited in the open configuration; in the closed state: at least a portion of the sample is compressed by the two plates into a highly uniform layer, wherein the uniform thickness of the layer is limited by the plates and the light guide spacer in the sample area of ​​the plates; and

[0043] wherein, in a closed configuration, (a) at least one gasket in the sample contact area has its top surface in direct contact with one of the plates, and at least one gasket and the plate areas above and below it define a reference area, wherein the reference area is transparent to light within a specific wavelength range, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate are not occupied by a light-guiding gasket, thereby defining a sampling area that is transparent to light within a specific wavelength range.

[0044] 5. A device for analyzing hemoglobin in a sample, comprising:

[0045] A first board, a second board and a light guide spacer, wherein:

[0046] (v) the first plate and the second plate are configured to sandwich the sample in the thin layer;

[0047] A first board, a second board and a light guide spacer, wherein:

[0048] (i) the first plate and the second plate are configured to sandwich the sample in the thin layer;

[0049] (ii) the light-guiding spacer has a cylindrical shape and is sandwiched between two plates, wherein the head end and the tail end of the column directly contact one of the plates, so that there is no sample between the two ends of the column and the corresponding plate, wherein the light-guiding spacer is surrounded by the sample or in the vicinity of the sample, wherein the direct contact area and the average cross-section of the column are at least 1 um^2 (square micrometer) or greater, respectively;

[0050] The interval between the inner surfaces of the two plates is 200 μm or less.

[0051] 6. A sample analysis device, comprising:

[0052] Any one of the present device embodiments, a light source, a camera; and an adapter, wherein

[0053] (i) the light source is arranged to emit light within a certain wavelength range, and the light within the wavelength range can pass through the reference area;

[0054] (ii) The camera is configured to image the reference area and the sampling area.

[0055] (iii) The adapter is configured to position the device, the light source and the camera relative to each other so that light from the light source passes through the reference region and the sampling region and is imaged by the camera.

[0056] 7. The apparatus of any preceding apparatus embodiment, further comprising:

[0057] A processor is configured to process the image captured by the camera and determine a property of an analyte in the sample by comparing light transmission from the reference region and the sampling region.

[0058] 8. An apparatus as claimed in any preceding claim, wherein the camera and the processor are part of a single mobile device.

[0059] 9. An apparatus according to any preceding claim, wherein the light source and the processor are part of a single mobile device.

[0060] 10. A method for sample analysis using transmitted light, comprising the following steps:

[0061] (a) a device having any of the preceding apparatus embodiments;

[0062] (b) depositing a sample in an open configuration of the device, wherein the sample may contain an analyte;

[0063] (c) placing the device into a closed configuration;

[0064] (d) constructing a light source having a certain wavelength to pass through the reference region of the device;

[0065] (e) constructing an imager for imaging the reference region and the sampling region of the device;

[0066] (f) constructing an adapter configured to position the device, the light source, and the camera relative to each other so that light from the light source passes through the reference region and the sampling region and is imaged by the camera;

[0067] (g) Determine the nature of the analyte by comparing the light transmittance from the sample area and the reference area.

[0068] Incorporated by Reference

[0069] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Those skilled in the art will appreciate that the drawings described below are for illustrative purposes only. In some figures, the drawings are drawn to scale. In the figures providing experimental data points, the lines connecting the data points are only used to guide the data view, without other means. For clarity, some elements are enlarged when shown in the drawings. It should be noted that these drawings are not intended to show these elements strictly to scale. The size of the elements should be delineated from the description provided herein and incorporated herein by reference. The drawings are not intended to limit the scope of the present invention in any way.

[0072] Figure 1A A cross-sectional view of one embodiment of a sample holder referred to as an OAC (e.g., optical analysis card) is shown, which is used to analyze an analyte in a sample (e.g., hemoglobin in a blood sample) by using light transmission, comprising: a first plate, a second plate, and a light guide gasket (LGS); wherein the LGS has a columnar shape, is sandwiched between the two plates, and each end of the column directly contacts one of the two plates forming an LGS-plate contact area, and is configured to allow light to be transmitted from the first plate through the LGS and move to the second plate without passing through the sample.

[0073] Figure 1B A cross-sectional view of one embodiment of a sample holder, referred to as an OAC (e.g., optical analysis card), for optically analyzing an analyte in a sample (e.g., hemoglobin in a blood sample) using optical transmission is shown, comprising: a first plate, a second plate and a light guide spacer (LGS), wherein the two plates are movable relative to each other and the top of the support is flat.

[0074] Figure 1C is an illustration of a CROF (Compressed Regulated Open Flow) embodiment. FIG. (a) shows a first plate and a second plate, wherein the first plate has a gasket. FIG. (b) shows depositing a sample on a first plate (shown), a second plate (not shown), or both (not shown) in an open configuration. FIG. (c) shows (i) using two plates to spread the sample (sample flow between the plates) and reduce sample thickness, and (ii) using gaskets and plates to regulate sample thickness in a closed configuration. The inner surface of each plate may have one or more bonding areas and / or storage areas (not shown).

[0075] Figure 2A A cross-sectional view showing one embodiment of a sample holder having a first plate, a second plate, and an LGS, a sample in the sample holder, positions of a sample region and a reference region, and incident and transmitted light in the sample region and the reference region

[0076] Figure 2B A perspective view of one embodiment of a sample holder having a first plate, a second plate, and an LGS is shown, samples in the sample holder, and the locations of sampling areas and reference areas.

[0077] Figure 3 A top view of one embodiment of a sample holder with a first plate, a second plate, and an LGS is shown, along with the location of the LGS, the sampling area, the reference area, and exemplary locations of the edges of the reference area and the sample area. Note that these edges are detected during the imaging process.

[0078] Figure 4The molar extinction coefficients of oxygenated hemoglobin [HbO2] and deoxygenated hemoglobin [Hb] at wavelengths of 200 nm to 1000 nm are shown.

[0079] Figure 5 Shown is the optical setup for measuring hemoglobin in a QMAX card.

[0080] Figure 6 An exemplary hemoglobin measurement in a QMAX card performed by an iPhone is shown.

[0081] Figure 7 An exemplary QMAX hemoglobin measurement compared to the gold standard (Abbott Emerald hemocytometer) is shown.

[0082] Figure 8 An exemplary hemoglobin measurement from a QMAX card taken by an iPhone is shown.

[0083] Fig. 9 An exemplary method for selecting sampling regions and reference regions is shown. Detailed description

[0085] The following detailed description illustrates certain embodiments of the present invention by way of example and not limitation. The section headings and any subheadings used herein, if any, are for organizational purposes only and should not be construed as limiting the subject matter described in any way. The content under a section heading and / or subheading is not limited to the section heading and / or subheading, but applies to the entire description of the present invention.

[0086] The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present claims are not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0087] definition

[0088] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present teachings, some exemplary methods and materials are now described.

[0089] The term "light guide spacer" or "LGS" may refer to a support having one end that directly contacts a first plate and another end that directly contacts another plate during light transmission measurement of the sample. The second plate, in some embodiments, sandwiches the sample between the first plate and the second plate. In some embodiments, the dimensions of the optical indicators and the support are predetermined and known. In some embodiments, the LGS is made of the same material as one or both plates. In some embodiments, the LGS is bonded, molded, embossed, or otherwise attached to one or both plates.

[0090] The term "no significant amount of sample" may refer to an amount of sample that is insignificant to light transmission measurement of the sample when measurement is performed in an area having two plates and the sample.

[0091] The term "LGS-plate contact area" may refer to the area of ​​both ends of the LGS (having a cylindrical shape) that is in direct contact with one of the plates. In some embodiments, the LGS and one plate are made of one piece of material, then the LGS-plate contact area of ​​the end of the LGS connected to the plate is the cross-section of the LGS. In some embodiments, the LDG and both plates are made of one piece of material, then the LGS-plate contact area of ​​both ends of the LGS is the cross-section of the LGS.

[0092] The term “lateral cross-section of the LGS” may refer to a cross-section of the LGS parallel to the plates when the LGS is sandwiched between two plates.

[0093] The term “LGS contact area or LGS cross-section is larger than the wavelength of light” may mean that the LGS contact area or LGS cross-section is larger than or equal to the area of ​​a disk having a diameter of the wavelength of light.

[0094] The term "OTSA" refers to Light Transmission Sample Analysis, which measures the optical density of a thin sample layer by light transmission.

[0095] The terms "SR region" or "a pair of SR regions" are interchangeable and may refer to a sampling region and a set of corresponding reference regions, where the OD of the thin sample layer is determined by taking the ratio of the light intensity transmitted through the sample region and the reference region.

[0096] The term "reference region" of the OAC device may refer to a region where light of a specific wavelength and polarization passes through the first plate, the light-guiding spacer, and the second plate, wherein the light-guiding spacer is in direct contact with the first and second plates. The term "reference region" of the OAC device may refer to a region where the light-guiding spacer is sandwiched between the two plates and is in direct contact with each plate, respectively, wherein, in the reference region, the probe light sequentially passes through the first plate, the light-guiding spacer, and the second plate without passing through the sample.

[0097] The term “sampling region” of an OAC device refers to a region on the device where light of a specific wavelength and polarization can pass through the reference region, through the sample between two plates, and the second plate without passing through the light-guiding spacer.

[0098] The term "sampling region" of an OAC device refers to the region of the device where the upper sample is located between the two plates and there is no LGS in this region. That is, in the sampling region, the probe light sequentially passes through the first plate, the sample between the two plates, and the second plate without encountering the LGS.

[0099] The term “distance between the sampling region and the reference region” of the OAC device may refer to the shortest interval between a boundary of the reference region and a boundary of the sampling region.

[0100] The terms "imager" and "camera" are used interchangeably.

[0101] The terms pillar, an LSG, a column or an object "inside the sample" mean that the side walls of these objects are surrounded by the sample.

[0102] The term "embossing" refers to the integral fastening of the shim and the plate by impressing (eg, embossing) a piece of material to form the shim on the surface of the plate. The material may be a single layer of material or multiple layers of material.

[0103] The term "etched" means that the pad and board are integrally secured by etching a piece of material to form the pad on the board surface. The material can be a single layer of material or multiple layers of material.

[0104] The term "fused to" means that the gasket and the plate are integrally fixed by bonding them together, original materials for the gasket and the plate are fused to each other, and there is a clear material boundary fusion between the two materials afterwards.

[0105] The term "bonded to" means that the gasket and the plate are integrally fixed by bonding the gasket and the plate through adhesion.

[0106] The term "attached to" means that the gasket and the plate are connected together.

[0107] The terms "CROF card (or card)", "COF card", "QMAX card", "Q card", "CROF device", "COF device", "QMAX device", "CROF board", "COF board" and "QMAX board" are interchangeable, but in some embodiments, the COF card does not include a gasket. And the terms refer to a device that includes a first board and a second board that can be relatively moved to different configurations (including an open configuration and a closed configuration) and includes a gasket (except for certain embodiments of the COF card) to adjust the spacing between the boards. The term "X board" can refer to one of the two boards in the CROF card, where the gasket is fixed to the board. Further description of the COF card, CROF card and X board is provided in the provisional application serial number. The entire content of U.S. Patent No. 62 / 456,065 filed on February 7, 2017 is incorporated herein.

[0108] In the QMAX process, the term "open configuration" of two plates refers to a configuration in which the two plates are partially or completely separated and the spacing between the two plates is not limited by spacers.

[0109] In the QMAX process, the "closed configuration" of two plates refers to the following configuration: the two plates are opposite to each other, the spacer and a certain volume of the sample are located between the two plates, and the relative spacing between the two plates is such that the thickness of the relevant volume of the sample is adjusted by the plates and the spacer, wherein the relevant volume is at least a portion of the entire volume of the sample.

[0110] In the QMAX process, the term "sample thickness is regulated by the plate and the shim" means that for given conditions of the plate, sample, shim and plate compression method, the thickness of at least one orifice is certain. The thickness of the portion of the sample when the plate is in the closed configuration can be predetermined based on the characteristics of the shim and the plate.

[0111] The "inner surface" or "sample surface" of the plate in the QMAX card may refer to the surface of the plate that is in contact with the sample, while the other surface of the plate (not in contact with the sample) is referred to as the "outer surface."

[0112] Unless otherwise specified, in the QMAX process, the "height" or "thickness" of an object may refer to the dimension of the object in the direction perpendicular to the surface of the board. For example, the shim height is the dimension of the shim in the direction perpendicular to the surface of the board, and the shim height and shim thickness refer to the same thing.

[0113] Unless otherwise specified, in the QMAX process, the term "area of ​​an object" refers to the area of ​​the object parallel to the surface of the board. For example, the pad area is the area of ​​the pad parallel to the surface of the board.

[0114] The term QMAX card may refer to a device that performs a QMAX (eg, CROF) process on a sample and may or may not have a hinge connecting the two plates.

[0115] The terms "QMAX card with hinge" and "QMAX card" are interchangeable.

[0116] The terms "self-holding angle", "automatic angle holding" or "automatic rotation angle" refer to the property of a hinge that essentially maintains the angle between two panels after an external force moves the panels into that angle from an initial angle and then removes the panels from that angle.

[0117] The terms "the shims have a predetermined height" and "the shims have a predetermined distance between shims" respectively mean that the values ​​of the shim height and the distance between shims are known before the QMAX process. If the values ​​of the shim height and the distance between shims are not known before the QMAX process, they are not predetermined. For example, in the case where beads are sprayed on the board as shims, the distance of the shims is not predetermined in the case where the beads fall at random positions on the board. Another example where the distance between shims is not predetermined is that the shims move during the QMAX process.

[0118] During the QMAX process, the term "shims are secured to their respective plates" means that the shims are secured to a certain position on the plates and remain attached to that position (i.e., the position of the shims on the respective plates does not change) during the QMAX process. An example of "shims are secured to their respective plates" is that the shims are integrally made of one material of the plates and the position of the shims relative to the plate surface does not change during the QMAX process. An example of "shims are not secured to their respective plates" is that the shims are bonded to the plates by an adhesive, but the adhesive fails to secure the shims to their original position on the plate surface and the shims move away from their original position on the plate surface during use of the plate during the QMAX process.

[0119] It will be apparent to those skilled in the art after reading the present invention that each individual embodiment described and illustrated herein has discrete components and features that can be easily separated or combined with the features of any of the other several embodiments without departing from the scope or spirit of the teachings of the present invention. Any narrated method can be performed in the order of events or any other order that is logically possible. It will be understood by those skilled in the art that the application of the present invention is not limited to the construction details, arrangement of components, category selection, weights, predetermined signal limits, or the steps set forth in the description or drawings herein. The present invention can have other embodiments and can be practiced or executed in many different ways.

[0120] Principles and some examples

[0121] One object of the invention relates to a device and a method for improving the light transmission analysis of a thin layer of a sample sandwiched between two plates, in particular for generating a reference signal that can improve the optical analysis, and for analyzing an analyte in a sample, such as hemoglobin in a blood sample.

[0122] Certain biological or chemical properties of a sample can be determined by measuring the absorption coefficient α of a thin sample layer in an experiment of light transmission through the sample layer. s Using Beer-Lambert's law, the light absorption coefficient of the thin sample layer, α s The incident light intensity (i.e. the light incident on the sample) I i and the transmitted light intensity (i.e. the light passing through the sample) I t related,:

[0123]

[0124] The length of the sample layer (i.e. thickness) is the light density passing through the sample layer. The light absorption coefficient of the thin sample layer is α s can be related to the properties of the sample. Therefore, using the Beer-Lambert law, the properties of the sample can be determined by measuring the OD of the sample layer.

[0125] However, in practice, it is difficult to directly measure the intensity of incident light (i.e. light that directly hits the sample layer) and transmitted light (i.e. light that directly transmits through the sample). Usually, the total light transmittance through the sample and the sample holder is measured in experiments (this is because thin layer samples usually require a sample holder for measurement, and the light being measured also passes through the sample holder). Therefore, it is necessary to separate / determine the sample's OD from the total transmittance.

[0126] According to the present invention, a specific sample holder called OAC (i.e., Optical Analysis Card) is provided, and the optical density of the material is determined by taking the ratio of the intensities of two transmitted lights: one is the light beam passing through the sampling area of ​​the sample holder, and the other is the light transmitted through the reference area of ​​the sample holder, thereby determining the OD of the sample without directly measuring the incident light.

[0127] Sample rack

[0128] According to the present invention, as shown in FIGS. 1 and 2 , an embodiment of a sample holder called an OAC (i.e., optical analysis card) for analyzing an analyte in a sample (e.g., hemoglobin in a blood sample) by light transmission using light comprises:

[0129] The first board, the second board, the light guide spacer (LGS), the sampling area and the reference area, where:

[0130] (i) the first plate and the second plate are configured to sandwich a sample for optical transmission analysis by light, sandwiching it in a thin layer between the plates, and each plate has a sample contact area on its inner surface for contact with the sample;

[0131] (ii) a light guide spacer (LGS) having a cylindrical shape, sandwiched between the two plates, with each end of the column directly contacting one of the plates forming the LGS plate contact area, and configured to allow light to be transmitted from the first plate through the LGS to the second plate without passing through the sample,

[0132] (iii) a sampling region is a region where light can sequentially pass through the first plate, the sample, and the second plate, wherein the sampling region does not have an LGS; and

[0133] (iv) the reference area is the area where light passes through the first plate, the light guide spacer, and the second plate in sequence without passing through the sample;

[0134] Among them, the LGS contact area and the LGS cross section are larger than the wavelength of light,

[0135] wherein the light guide spacer is surrounded by the sample or is surrounded by the sample. and

[0136] Here, the thickness of the sample in the sampling area is 500 μm or less.

[0137] At least a portion of the plate in the reference area and the sampling area is light transmissive.

[0138] According to the present invention, as shown in Figures 1 and 2, a sample holder called OAC (i.e., optical analysis card) has at least a "sampling area" and a "reference area", and the light absorption coefficient of the sample layer is determined by taking the ratio of light transmitted through the sampling area to light transmitted through the reference area.

[0139] In some embodiments, the sample holder (also referred to as a device) further comprises a plurality of light directing spacers having a substantially uniform height and wherein at least one of the light directing spacers is within the sample contact region.

[0140] In some embodiments, the first plate and the second plate are fixed by LGS ( Figure 1A ). In some embodiments, Figure 1B As shown, the first plate and the second plate can be moved relative to each other into different configurations including an open configuration and a closed configuration. In the open configuration, the plates are separated by a portion and the sample is deposited. In the closed configuration, the first plate and the second plate are respectively in contact with the flat end of the LGS.

[0141] In some embodiments, the first and second plates in the sample area and the reference have uniform thickness and are light transmissive.

[0142] The material of the plate is plastic, glass or other materials described in the present invention.

[0143] In some embodiments, additional shims are used to adjust the spacing between the first plate and the second plate, thereby adjusting the sample thickness.

[0144] Sample OD measurement method

[0145] According to the present invention, the properties of a sample are determined by measuring the OD of a thin layer of the sample, wherein the OD is determined by the ratio of light transmitted through a sampling region of the OAC to light transmitted through a reference region of the OAC.

[0146] In some embodiments, an image of a sample in a sample holder is captured by a camera and analyzed. (e.g. Figure 5 )

[0147] In some embodiments, the wavelength of the light is in the range of 500 nm to 1200 nm, 200 nm to 3000 nm, 3000 nm to 30,000 nm, or 100 nm to 200 nm.

[0148] A) Light absorption through the sample, which is determined by the light transmittance in the sample and reference regions. For light with incident light intensity I0, the transmitted light intensity I through the sample is determined by the Beer-Lambert law. s Given by:

[0149]

[0150] Where, is the extinction coefficient of the sample (e.g., hemoglobin), c is the average concentration of the sample (e.g., hemoglobin), and L is the optical path length through the sample. (ε is in cm -1 / M, c in M, L in cm), OD s It can express the optical density of the sample.

[0151] For the incident light intensity I0, the light travel length is L r The transmitted light intensity after the light guide spacer, I r Yes, using the Beer-Lambert law, it is given by:

[0152]

[0153] In the formula, α r is the absorption coefficient of the light guide spacer, L is the optical path length through the sample, OD s Can refer to the light density passing through the light guide spacer.

[0154] Subtracting the first equation from the second gives:

[0155]

[0156] According to the present invention, the above equation shows that the absorption coefficient of the sample layer can be determined by obtaining the ratio of the transmitted light through the sampling area to the transmitted light through the reference area without measuring the incident light (assuming that the incident light in the two areas is exactly the same).

[0157] Use spacers to create a uniform thin layer of sample.

[0158] Fig. 1 is a diagram of a CROF (Compressed Regulated Open Flow) embodiment. Fig. (a) shows a first plate and a second plate, wherein the first plate has a gasket. Fig. (b) shows that a sample is deposited on a first plate (shown), a second plate (not shown) or both (not shown) in an open configuration. Fig. (c) shows (i) using two plates to spread the sample (sample flow between the plates) and reduce the sample thickness, and (ii) using a gasket and a plate to regulate the sample thickness in a closed configuration. The inner surface of each plate may have one or more binding sites and / or storage sites (not shown).

[0159] B) Two types of hemoglobin

[0160] There are two types of hemoglobin in the blood. Oxygenated hemoglobin [HbO2] is the form of hemoglobin that is bound to oxygen, while deoxygenated hemoglobin [Hb] is the form of hemoglobin that is not bound to oxygen. Normally, oxygenated hemoglobin [HbO2] is about 75% in veins and about 90% in arteries.

[0161] Total hemoglobin concentration = [HbO2] + [Hb].

[0162] like Figure 4 As shown, the two hemoglobins may have different extinction coefficients (i.e. light absorption) at different wavelengths. Therefore, by measuring the light absorption of the blood in different wavelength ranges, the concentrations of [HbO2] and [Hb] in the blood can be determined separately.

[0163] C) Analyze the light transmission sample by comparing the light transmission from the sample area and the reference area

[0164] In accordance with the present invention, the absorption of light (and optical density ("OD")) through a thin sample layer is measured by comparing the light transmittance from the sample area to that from a reference area.

[0165] In some cases, it is necessary to compare the light transmittance from a sample area to a reference area.

[0166] D) Improved analysis of light transmission samples

[0167] In many real-world measurement situations, there are a number of imperfections that can significantly reduce the accuracy of the OD measurement. For example, the sample in the sample holder and / or the sample holder itself may have non-uniform thickness. There are imperfections in the sample or sample holder, such as bubbles, dust, or other imperfections that may cause the optical transmission to differ from that through an ideal sample (i.e., a perfect sample). The light intensity may not be consistent across the measurement area.

[0168] The present invention has multiple ways to reduce errors in optical transmission sample analysis (OTSA) due to defects. According to the present invention, in order to improve OD measurement accuracy, the following features, devices and methods below (i.e., in Section 1.4 and its subsections) are used alone or in combination.

[0169] I. Reduce light scattering through LGS sidewalls and / or LGS-Plate interface

[0170] According to the present invention, in one embodiment of the OD measurement method, the OD measurement method measures the light intensity of the sample area and the reference area, and then takes the ratio of the two intensities. If the light passing through the reference area is strongly scattered from (a) the LGS sidewall or (b) the LGS, or the light emitted from the sample area is significantly scattered from the nearby LGS sidewall, then the measurement accuracy may be significantly reduced.

[0171] To reduce the effect of light scattered by the LGS sidewalls on the light from the reference area, the edge of the reference area used to determine the OD should be a certain distance from the LGS sidewalls. Since the reference area cannot be smaller than the wavelength of light without significant light diffraction, to reduce the effect of light scattered by the LGS sidewalls on the light emitted from the reference area, at least the cross-section of the LGS should be larger than the wavelength of light.

[0172] In some embodiments, the edge of the reference area used to determine the OD is a distance from the LGS sidewall.

[0173] In some embodiments, the cross-section of the LGS should be larger than the wavelength of light and at a distance from the LGS sidewall at the edge of the reference area used to determine the OD.

[0174] Similar to the light from the reference area, to reduce the effect of light scattering on the light from the sampling area, the edge of the sampling area should be a certain distance away from the LGS side wall.

[0175] In some embodiments, the edge of the sampling area used to determine the OD is a distance away from the LGS sidewall.

[0176] In some embodiments, an edge of a reference area for determining OD is a distance from a LGS sidewall, and an edge of a sampling area for determining OD is a distance from a LGS sidewall.

[0177] In some embodiments, the cross-section of the LGS should be larger than the wavelength of light, the edge of the reference area for OD determination is at a distance from the LGS sidewall, and the edge of the sampling area for OD determination is at a distance from the LGS sidewall.

[0178] II. The area of ​​the reference area and the sampling area, and the distance between them

[0179] When determining the OD of a sample by obtaining the ratio of the light intensity passing through the sample area and the light intensity passing through the reference area, it is assumed that the incident light in each area has the same intensity, or that the thicknesses of the first plate and the second plate of the sample are the same or known in the sampling area and the reference area, respectively. However, in many actual optical systems, the above assumptions do not hold true, which leads to uncertainty (i.e., error) in determining the OD. For example, in reality, the intensity of the incident light used for sample light transmission measurement is not uniform, especially when the illumination area is large. The thicknesses of the first plate, the second plate, and the sample are different or unknown in the sampling area and the reference area, respectively, and each can have significant variations.

[0180] According to the present invention, one method to reduce the error is to limit the area of ​​the sampling area and the reference area used to determine the OD of the sample, or to make the distance between the sampling area and the reference area smaller while avoiding LGS side wall scattered light, or both.

[0181] In some embodiments, the area of ​​the sampling region and the distance between the sampling region and the reference region are a combination of the above two paragraphs.

[0182] III. Multiple pairs of sampling and reference areas

[0183] Using a pair of sample area and reference area will lead to large errors. This is due to the following reasons: (i) Since the spatial variations of the thickness of the first plate, the second plate and the sample are random, only a pair of sample area and reference area may not represent most of the samples; (ii) Since the number of optical defects and their locations are also random, these optical defects will occur at the location of the sample area and / or the reference area, making the sample area and reference area pair unusable in OTSA.

[0184] To solve these problems, according to the present invention, multiple pairs of SR regions are used.

[0185] In some embodiments, the OAC includes multiple pairs of SR regions, wherein the distance between the centers of two adjacent SR regions is substantially periodic or aperiodic.

[0186] According to the present invention, reagents for facilitating testing are deposited on the inner surface of the OAC plate, including but not limited to staining reagents, surfactants, antibodies, proteins and nucleic acids.

[0187] Light guide (eg spacer)

[0188] In some embodiments, the light-guiding spacers are substantially periodic and predetermined in the distance between spacers.

[0189] Height of light guide

[0190] In some embodiments, the height of the light guide spacer is 1 μm, 2 μm, 5 μm, 10 μm, 30 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1,000 μm, 2,000 μm, 5,000 μm, 10,000 μm, or a range between two values.

[0191] Periodicity of light guide

[0192] In some embodiments, the shims are arranged in a periodic array of 1 μm, 2 μm, 5 μm, 10 μm, 30 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1,000 μm, 2,000 μm, 5,000 μm, 10,000 μm, or a range between two values.

[0193] Light Guide Spacer Geometry (LGS)

[0194] In some embodiments, the LGS has a cylindrical shape with substantially flat ends. In some embodiments, one or both ends of the LGS are secured to one or both of the boards by bonding, fusing, making from a single piece, or other methods of attaching the LGS to the board.

[0195] In some embodiments, the shape of the cross-section of the LGS includes, but is not limited to, a circle, a rectangle, a square, a triangle, a polygon, a letter, a number, or a combination thereof.

[0196] In some embodiments, the average cross-section (LGS) of each light guide spacer is 10μm^2 (square microns), 20μm^2, 30μm^2, 50μm^2, 100μm^2, 150μm^2, 200μm^2, 300μm^2, 500μm^2, 1000μm^2, 2000μm^2, 5000μm^2, 10,000μm^2, 30,000μm^2, 100,000μm^2, 200,000μm^2, 500,000μm^2, 1mm^2, 2mm^2, 5mm^2, 10mm^2, 50mm^2, or a range between two values.

[0197] In some preferred embodiments, the average cross-section of each light guide gasket is 1 μm^2 (square micrometer), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2, 10,000 μm^2, 30,000 μm^2, 100,000 μm^2, 200,000 μm^2 or a range between two values.

[0198] In certain preferred embodiments, the average cross-sectional guiding interval of each light is 1 μm^2 (square micrometer), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2, 10,000 μm^2, 30,000 μm^2 or a range between two values.

[0199] In certain preferred embodiments, the average cross-section of each light guide gasket is 1 μm^2 (square micrometers), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2 or in a range between either of the two values.

[0200] In some embodiments, the average cross-section of each light-guiding gasket is 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, larger than the wavelength of light passing through the reference area.

[0201] In certain preferred embodiments, the average cross-section of each light-guiding spacer is 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, or a range between two values, larger than the wavelength of light passing through the reference area.

[0202] Reference area geometry

[0203] shape

[0204] In some embodiments, the reference area is smaller than the smallest transverse cross-sectional dimension of the light guide rod. One advantage is that light scattering off the light guide sidewalls to affect the reference signal is avoided or reduced.

[0205] In some embodiments, the minimum distance between the edge of the light guide spacer and the edge of the reference area is 1 μm (micrometer), 2 μm, 3 μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1000 μm, or a range between two values.

[0206] In some preferred embodiments, the minimum distance between the edge of the light guide gasket and the edge of the reference area is 1 μm (micrometer), 2 μm, 3 μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, 100 μm, 200 μm or a range between two values.

[0207] In certain preferred embodiments, the minimum distance between the edge of the light guide spacer and the edge of the reference area is 1 μm (micrometer), 2 μm, 3 μm, 5 μm, 10 μm, 20 μm, 30 μm, 50 μm, or a range between two values.

[0208] In certain preferred embodiments, the minimum distance between the edge of the light-guiding gasket and the edge of the reference area is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, the wavelength passing through the reference area.

[0209] In certain preferred embodiments, the minimum distance between the edge of the light-guiding gasket and the edge of the reference area is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, the wavelength passing through the sampling area.

[0210] The ratio of the area of ​​the reference region to the area of ​​the light guide gasket is 3 / 10, 2 / 5, 1 / 2, 3 / 5, 7 / 10, 4 / 5, or a range between two values.

[0211] Sampling area geometry

[0212] In some embodiments, the edge of the sampling area is at a certain distance from the edge of the light guide rod. One advantage is to avoid or reduce light scattering from the side wall of the light guide rod to affect the reference signal.

[0213] In some preferred embodiments, the area of ​​the sampling region is 3 / 5, 7 / 10, 4 / 5, 9 / 10, 1, 11 / 10, 6 / 5, 13 / 10, 7 / 5, 3 / 2, or a range between two values ​​of the spacer interval.

[0214] In some preferred embodiments, the distance between the edge of the sampling area and the edge of the light guide gasket is 1 / 5, 3 / 10, 2 / 5, 1 / 2, 3 / 5, 7 / 10, 4 / 5, 9 / 10, 1 or a range between any two values.

[0215] In some preferred embodiments, the distance between the edge of the sampling area and the edge of the light-guiding gasket is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, of the wavelength passing through the reference area.

[0216] In some preferred embodiments, the distance between the edge of the sampling area and the edge of the light-guiding gasket is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, of the wavelength passing through the sampling area.

[0217] The distance between the sampling area and the reference area

[0218] In some embodiments, the distance between the edge of the sampling area and the reference area is 1 μm (micrometer), 2 μm, 3 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 100 μm, 200 μm, 500 μm, 1000 μm or a range between two values.

[0219] In certain preferred embodiments, the distance between the edge of the sampling area and the reference area is 30 μm (micrometers) to 50 μm, 20 μm to 60 μm, 10 μm to 70 μm, 5 μm to 75 μm, or between any two values ​​within this range.

[0220] In certain preferred embodiments, the distance between the edge of the sampling area and the reference area is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, of the wavelength passing through the reference area.

[0221] In some embodiments, the distance between the edge of the sampling area and the reference area is 2 / 5, 1 / 2, 3 / 5, 7 / 10, 4 / 5, 9 / 10, 1, 11 / 10, 6 / 5, 13 / 10, 7 / 5, 3 / 2, 8 / 5, 17 / 10, or in a range between two values.

[0222] In some embodiments, the distance between the edge of the sampling area and the reference area is 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, greater than the wavelength of light passing through the reference area.

[0223] In some embodiments, the distance between the edge of the sampling area and the reference area is greater than 1 times, 2 times, 3 times, 5 times, 10 times, 20 times, 50 times, 100 times, 200 times, 500 times, 1000 times, 5000 times, or a range between two values, of the wavelength passing through the sampling area.

[0224] Lysis reagent coating of iMOST-HgB device

[0225] In some embodiments, a surfactant is coated on the plates and dissolved into the blood to achieve uniform distribution of red blood cells in the device, where the coating can be on the first plate or the second plate or both.

[0226] In some embodiments, a surfactant is coated on the plates and lysed in the blood to lyse red blood cells in the device, wherein the coating can be on the first plate or the second plate or both.

[0227] In some embodiments, the surfactants coated in the device include, but are not limited to, zwitterions, ASB-14, ASB-16, CHAPS, cationic surfactants NN-[tris(hydroxymethyl)methyl]-N-alkyl-N,N-dimethylammonium chloride (IIa), llb, llc, lld, CTAC, Tween 20, Tween 40, Tween 60, Tween 80, sodium lauryl sulfate (SLS), ammonium lauryl sulfate, CTAB, sodium lauryl ether sulfate (SLES), sodium millesulfate, docusate, perfluorooctane sulfonate, alkyl aryl ether phosphate, alkyl ether phosphate, CTAB, cetylpyridinium chloride (CPC), benzene BAC, BZT, DIMETHYL DICHLORIDE, DOTAB, COCAMIDOPROPYL HYDROXYSULFONIC ACID, COCAMIDOPROPYL BETAINE, ETHOXYLATES, OCTAETHYLENE GLYCOL DODECYL ETHER, PENTAETHYLENE GLYCOL DODECYL ETHER, NONADYL OL, TRITON X-100, POLYETHOXYLATED TAUGHTAMINE, COCAMIDOETHANOLAMINE, COCAMIDIODIETHANOLAMINE, POLOXAMMER, GLYCERYL MONOSTEARATE, GLYCERYL MONOLAURATE, BITANNUM MONOLAURATE, SORBIAN MONOSTEARATE, SORBIAN TRISTEARATE, DECYL GLUCOSIDE, LAURYL GLUCOSIDE, OCTYL GLUCOSIDE, LAURYLDIMETHYLENE OXIDE, DIMETHYL SULFOXIDE, PHOSPHONIUM OXIDE.

[0228] In some embodiments, agents that cause red blood cell lysis in the device include but are not limited to Pluronic F-127, Cremophor EL, Pluronic F-68, Myrj 52, Brij 35, sodium oleate, sodium dodecyl sulfate, Tween 20, Tween 40, Tween 60, Tween 80, SLS, CTAB, CTAC, tamoxifen, saponin, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, lactic acid, ABS-14, ABS-16, antimalarial drugs (quinine compounds), arsenic, dapsone, metals (chromium / chromates, platinum salts, nickel compounds, copper, lead, cisplatin), nitrites, nitrofurantoin, penicillin, benzopyridine (pyridine), rho immune globulin, ribavirin, sulfonamides, sulfones.

[0229] In some embodiments, the anticoagulant coated in the device includes, but is not limited to, EDTA, such as dipotassium ethylenediaminetetraacetic acid (K2EDTA), tripotassium ethylenediaminetetraacetic acid (K3EDTA), coumarins (vitamin K antagonists), warfarin (coumarin), acenocoumarol, phenprocoumon, pyrimidine, phenindione, heparin, fondaparinux and idroparinux, dabigatran, rivaroxaban, apixaban, edoxaban, betrixaban, NOACs, hirudin, lepirudin, bivalirudin, argatroban, dabigatran, batroxobin, hirudin, vitamin E, sodium citrate, acid citrate dextrose, oxalates such as fluoride oxalate, citicoline, desirudin, enoxaparin.

[0230] In some embodiments, to achieve uniform distribution of red blood cells in the device, zwittergents were coated on the plate at the following concentrations: 3 ng / mm 2 ,5ng / mm 2 ,8ng / mm 2 ,12ng / mm 2 ,15ng / mm 2 ,25ng / mm 2 ,35ng / mm 2 ,50ng / mm 2 ,80ng / mm 2 ,100ng / mm 2 Or any range between two values.

[0231] In some embodiments, to lyse red blood cells in the device, zwittergent is added at 100 ng / mm 2 ,120ng / mm 2 ,150ng / mm 2 ,180ng / mm 2 ,200ng / mm 2 ,300ng / mm 2 ,400ng / mm2 ,500ng / mm 2 ,800ng / mm 2 ,1000ng / mm 2 Or coated on the board at a preferred area concentration in any range between the two values.

[0232] In some embodiments, to achieve uniform distribution of red blood cells in the device, the amphoteric bactericidal agent is coated on the plate, and the preferred final concentration in the blood is 0.05 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 1.0 mg / mL, 2 mg / mL or a range between two values.

[0233] In some embodiments, in order to lyse red blood cells in the device, Zwittergent is applied to the plate, and the preferred final concentration in the blood is 2 mg / mL, 3 mg / mL, 4 mg / mL, 5 mg / mL, 6 mg / mL. , 7 mg / mL, 9 mg / mL, 10 mg / mL, 15 mg / mL, 25 mg / mL, 50 mg / mL, or a range between two values.

[0234] Measuring range of iMOST-HgB devices

[0235] In some embodiments, the hemoglobin measurement range of the device is between 0 g / dL and 40 g / dL.

[0236] In some embodiments, the preferred hemoglobin measurement range of the device is between 0 g / dL and 30 g / dL.

[0237] In some embodiments, the preferred hemoglobin measurement range of the device is between 5 g / dL and 26 g / dL.

[0238] Removal and compensation of scattering particles

[0239] In some cases, there are imperfections that can significantly reduce the accuracy of absorbance and / or hemoglobin measurements.

[0240] For example, the presence of interfering particles in the sample or sample holder includes but is not limited to particles that increase turbidity, light scattering particles, bubbles, dust or other particles that may have a different light transmittance than through a perfect sample (i.e., an ideal sample).

[0241] The present invention has several ways to reduce errors in optical transmission sample analysis (OTSA) due to defects.

[0242] In some embodiments, multiple pairs of SR regions are used. For each pair of SR regions, the OD of the sample is determined by taking the ratio of the light intensity passing through the sample region and the light intensity passing through the reference region. For a given pair of SR regions, a quality measure of the SR region is calculated. If the quality measure is low, the pair of SR regions is excluded from the merging algorithm in the next stage. The pooling algorithm is to pool the OD of the sample across all pairs of SR regions. Various merging algorithms can be utilized, including but not limited to median, mean, maximum, minimum, k-means, etc.

[0243] In some embodiments, defective areas are removed or excluded from the image prior to light intensity analysis.

[0244] In some embodiments, defective regions having borders are removed or excluded from the image prior to light intensity analysis, wherein the border size is between 1 μm and 50 μm; and wherein a preferred border size is between 5 μm and 20 μm.

[0245] Measurement with / without scanning

[0246] In certain embodiments, a location on the device is measured for analysis, particularly for hemoglobin.

[0247] QMAX System

[0248] A) QMAX card

[0249] Details of the QMAX card are described in various publications, including International Application No. PCT / US2016 / 046437 (Essenlix Docket No. ESSN-028WO), which is hereby incorporated by reference for all purposes.

[0250] I. Tablet

[0251] In the present invention, generally, the plates of the CROF are made of any material that (i) can be used with shims to adjust the thickness of a portion or all of the volume of the sample, and (ii) does not have any material that would have a significant adverse effect on the sample, assay, or goal that the plate is intended to achieve. However, in certain embodiments, specific materials (and therefore their properties) are used for the plates to achieve certain goals.

[0252] In certain embodiments, the two panels have the same or different parameters for each of the following parameters: panel material, panel thickness, panel shape, panel area, panel flexibility, panel surface properties, and panel optical transparency.

[0253] (I) Plate material. The plate is made of a single material, a composite material, multiple materials, a multi-layer material, an alloy or a combination thereof. Each material of the plate is an inorganic material, an organic material or a mixture, wherein examples of the materials are given in the paragraphs of Mat-1 and Mat-2.

[0254] Mat-1: The inorganic materials of the sheet include but are not limited to glass, quartz, oxides, silicon dioxide, silicon nitride, helium oxide (HfO), aluminum oxide (AIO), semiconductors: (Si, GaAs, GaN, etc.), metals (such as gold, silver, copper, aluminum, Ti, Ni, etc.), ceramics or any combination thereof.

[0255] Mat-2: Organic materials used for the gasket include but are not limited to polymers (such as plastics) or amorphous organic materials. The polymer material for the gasket includes, but is not limited to, acrylate polymers, vinyl polymers, olefin polymers, cellulosic polymers, non-cellulosic polymers, polyester polymers, nylon, cyclic olefin copolymer (COC), polymethyl methacrylate (PMMA), polycarbonate (PC), cyclic olefin polymer (COP), liquid crystal polymer (LCP), polyamide (PA), polyethylene (PE), polyimide (PI), polypropylene (PP), polyphenylene ether (PPE), polystyrene (PS), polyoxymethylene (POM), polyetheretherketone (PEEK), polyethersulfone (PES), polyethylene phthalate (PET), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), polyvinylidene fluoride (PVDF), polybutylene terephthalate (PBT), fluorinated ethylene propylene (FEP), perfluoroalkoxyalkane (PFA), polydimethylsiloxane (PDMS), rubber or any combination thereof.

[0256] In some embodiments, the plates are each independently made of at least one of glass, plastic, ceramic, and metal. In some embodiments, each plate independently includes at least one of glass, plastic, ceramic, and metal.

[0257] In some embodiments, one plate is different from another plate in lateral area, thickness, shape, material or surface treatment. In some embodiments, one plate is the same as another plate in lateral area, thickness, shape, material or surface treatment.

[0258] The material of the panels is rigid, flexible or anything in between. Rigidity (eg, stiffness) or flexibility is relative to a given pressure for bringing the panels into a closed configuration.

[0259] In certain embodiments, the choice of a rigid or flexible plate is determined by the need to control the uniformity of sample thickness in a closed configuration.

[0260] In some embodiments, at least one of the two plates is transparent (to light). In some embodiments, at least a portion or portions of one or both plates are transparent. In some embodiments, the plates are opaque.

[0261] (ii) Plate Thickness. In certain embodiments, at least one plate has an average thickness of 2 nm or less, 10 nm or less, 100 nm or less, 500 nm or less, 1000 nm or less, 2 μm (micrometer) or less, 5 μm or less, 10 μm or less, 20 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 millimeter (millimeter) or less, 2 mm or less, 3 mm or less, or a range between any two values.

[0262] In certain embodiments, at least one plate has an average thickness of at most 3 mm (millimeter), at most 5 mm, at most 10 mm, at most 20 mm, at most 50 mm, at most 100 mm, at most 500 mm, or a range between any two values.

[0263] In some embodiments, the thickness of the plate is not uniform across the plate. Using different plate thicknesses at different locations can be used to control plate bending, folding, sample thickness adjustment, etc.

[0264] (iii) Shape and area of ​​the plate. In general, the plate can have any shape as long as the shape allows for compressed open flow of the sample and adjustment of the sample thickness. However, in certain embodiments, a specific shape may be advantageous. The shape of the plate can be circular, oval, rectangular, triangular, polygonal, annular, or any superposition of these shapes.

[0265] In some embodiments, the two plates can have the same size or shape, or different. The area of ​​the plate depends on the application. The area of ​​the plate is up to 1 square millimeter (square millimeter), up to 10 square millimeters, up to 100 square millimeters, up to 1 square millimeter (square centimeter), up to 5 square centimeters, up to 10 square centimeters, up to 100 square centimeters. Up to 500 square centimeters, up to 1000 square centimeters, up to 5000 square centimeters, up to 10,000 square centimeters, or more than 10,000 square centimeters, or any arrangement between two values. The shape of the plate can be rectangular, square, circular or other shapes.

[0266] In some embodiments, at least one of the panels is in the form of a strip (or strips) having a width, a thickness, and a length. The width is at most 0.1 centimeters (cm), at most 0.5 cm, at most 1 cm, at most 5 cm, at most 10 cm, at most 50 cm, at most 100 cm, at most 500 cm, at most 1000 cm, or a range between any two values. The length can be a desired length. The strip panels can be rolled into rolls.

[0267] (iv) Plate Surface Flatness. In many embodiments, the inner surface of the plate is flat or remarkably flat, planar. In certain embodiments, the two inner surfaces are parallel to each other in the closed configuration. A flat inner surface can facilitate quantification and / or control of sample thickness by simply using a predetermined shim height in the closed configuration. For a non-flat inner surface of the plate, not only does one need to know the shim height, but one also needs to know exactly the topology of the inner surface to quantify and / or control the sample thickness in the closed state. Knowing the surface topology requires additional measurements and / or corrections, which can be complex, time consuming, and expensive.

[0268] The flatness of the flat surface is relative to the final sample thickness (the final thickness is the thickness in the closed configuration) and is often characterized by the term "relative surface flatness," which is the ratio of the final sample thickness to the final sample thickness.

[0269] In some embodiments, the relative surface is less than 0.01%, 0.1%, less than 0.5%, less than 1%, less than 2%, less than 5%, less than 10%, less than 20%, less than 30%, less than 50%, less than 70%, less than 80%, less than 100%, or a range between any two of these values.

[0270] (v) Plate surface parallelism. In some embodiments, the two surfaces of the plate are substantially parallel to each other. In some embodiments, the two surfaces of the plate are not parallel to each other.

[0271] (vi) Flexibility of the plates. In some embodiments, the plates are flexible under the compression of the CROF process. In some embodiments, both plates are flexible under the compression of the CROF process. In some embodiments, one plate is rigid and the other plate is flexible under the compression of the CROF process. In some embodiments, both plates are rigid. In some embodiments, both plates are flexible, but have different flexibilities.

[0272] (vii) Transparency of the panels. In some embodiments, the panels are optically transparent. In some embodiments, both panels are optically transparent. In some embodiments, a panel is optically transparent and the other panel is opaque. In some embodiments, both panels are opaque. In some embodiments, both panels are optically transparent, but have different optical transmittances. The optical transparency of a panel may refer to a portion or the entire area of ​​the panel.

[0273] (ⅷ) Surface wettability. In certain embodiments, the inner surface of the plate wets (e.g., contact angle is less than 90 degrees) the sample, the transferred liquid, or both. In certain embodiments, both plates have inner surfaces that wet the sample, the transferred liquid, or both; and, the two surfaces have wettability. Have the same or different wettabilities. In certain embodiments, the plate has an inner surface that wets the sample, the transferred liquid, or both. The inner surface of the other plate does not wet (e.g., contact angle is equal to or greater than 90 degrees). Wetting of the inner surface of the plate can refer to a portion or the entire area of ​​the plate.

[0274] In certain embodiments, the inner surface of the plate has other nano- or micro-structures to control the lateral flow of the sample during CROF. The nano- or micro-structures include, but are not limited to, channels, pumps, etc. Nano- and micro-structures are also used to control the wetting properties of the inner surface.

[0275] II. Gasket

[0276] (i) Function of the gasket. In the present invention, the gasket is configured to have one or any combination of the following functions and characteristics: the gasket is configured to (1) control the thickness of the sample or the relevant volume of the sample together with the plate (preferably, the thickness control is precise, or uniform, or both within the relevant area); (2) allow the sample to have a compressed regulated open flow (CROF) on the plate surface; (3) not take up a significant surface area (volume) in a given sample area (volume); (4) reduce or increase the effect of particle or analyte sedimentation in the sample; (5) change and / or control the wettability of the inner surface of the plate; (6) determine the position, size ratio and / or information related to the plate, or (7) any combination of the above.

[0277] (ii) Shim structure and shape. In order to achieve the purpose of reducing and controlling sample thickness, in some embodiments, the shims are fixed to their respective plates. Generally, the shims can have any shape as long as the shims can adjust the sample thickness during the CROF process, but in order to achieve certain functions (such as better uniformity, degree of overshoot of pressing, etc.), certain shapes are preferably used.

[0278] A sheet is a single shim or a plurality of shims (e.g., an array). Certain embodiments of a plurality of shims are arrays of shims (e.g., columns) where the distances between shims are periodic or aperiodic, or are periodic or aperiodic in certain areas of the sheet, or have different distances in different areas of the sheet.

[0279] There are two types of shims: open shims and closed shims. An open shim is one that allows sample flow through the shim (e.g., the sample flows through and through the shim. For example, a column acts as a shim.), and a closed shim is one that blocks sample flow. (e.g., the sample cannot flow through the shim. For example, a ring shim, the sample is inside the ring.) Both types of shims use their height to adjust the final sample thickness in a closed configuration.

[0280] In some embodiments, the gasket is only an open gasket. In some embodiments, the gasket is only a closed gasket. In some embodiments, the gasket is a combination of an open gasket and a closed gasket.

[0281] The term "cylindrical spacer" means that the spacer has a cylindrical shape, and the cylindrical shape may refer to an object having a height and side shape that allows a sample to flow around it during compressed open flow.

[0282] In some embodiments, the lateral shape of the columnar spacer is a shape selected from (i) a circle, an ellipse, a rectangle, a triangle, a polygon, a ring, a star, a letter shape (e.g., an L shape, a C shape, a letter from A to Z), a number shape (e.g., a shape such as 0, 1, 2, 3, 4... to 9); (ii) a shape in the (i) group having at least one rounded corner; (iii) a shape in the (i) group having a serrated or rough edge; (iv) any superposition of (i), (ii) and (iii). For multiple spacers, different spacers may have different lateral shapes and sizes and different distances from adjacent spacers.

[0283] In some embodiments, the spacers can be and / or include columns, posts, beads, balls, and / or other suitable geometric shapes. The lateral shape and size of the spacers (e.g., transverse to the corresponding plate surface) can be anything, except that in some embodiments, there are the following restrictions: (i) the geometry of the spacers does not cause significant errors and quantities when measuring the sample thickness; or (ii) the geometry of the spacers does not prevent the sample from flowing out between the plates (e.g., it is not a closed form). However, in some embodiments, they require some spacers to be closed spacers to restrict sample flow.

[0284] In some embodiments, the shape of the gasket has rounded corners. For example, a rectangular gasket has one, several, or all chamfered corners (e.g., rounded instead of 90 degree angles). Rounded corners generally make the gasket easier to manufacture and, in some cases, less damaging to the biological material.

[0285] The sidewalls of the struts can be straight, curved, angled, or different shapes in different portions of the sidewalls. In some embodiments, the spacers are posts with various lateral shapes, sidewalls, and lateral area ratios of post height to post height. In a preferred embodiment, the spacers have a strut shape for allowing open flow.

[0286] (iii) Spacer material. In the present invention, the spacer is generally made of any material that can adjust the thickness of the relevant volume of the sample together with the two plates. In some embodiments, the material used for the spacer is different from the material used for the plates. In some embodiments, the material used for the space is at least partially the same as the material used for at least one of the plates.

[0287] The gasket is made of a single material, a composite material, multiple materials, multi-layer materials, alloys or a combination thereof. Each material used for the gasket is an inorganic material, an organic material or a mixture, wherein examples of materials are given in the paragraphs of Mat-1 and Mat-2. In a preferred embodiment, the gasket is made of the same material as the plate used in the CROF.

[0288] (iv) Mechanical strength and flexibility of the gasket. In some embodiments, the mechanical strength of the gasket is strong enough so that during compression and in the closed configuration of the panel, the height of the gasket is the same or significantly the same as the height of the panel in the open configuration. In some embodiments, the difference between the open configuration and the closed configuration of the gasket can be characterized and predetermined.

[0289] The material of the spacer is rigid, flexible or anything in between. Rigidity is relative to the applied pressure used to bring the panels into the closed configuration: if the space deforms by no more than 1% of its height under the pressure, the spacer material is considered rigid, otherwise it is considered flexible. When the spacer is made of a flexible material, the final sample thickness in the closed configuration can still be determined based on the pressing force and mechanical properties of the spacer.

[0290] (v) spacers in the sample. In order to achieve the desired sample thickness reduction and control, in particular to obtain good sample thickness uniformity, in certain embodiments, spacers are placed inside the sample or in the relevant volume of the sample. In certain embodiments, one or more spacers are present inside the sample or in the relevant volume of the sample, with appropriate spacing between the spacers. In certain embodiments, at least one spacer is inside the sample, at least two spacers are inside the sample or in the relevant volume of the sample, or at least "n" spacers are inside the sample or in the relevant volume of the sample, where "n" can be determined by sample thickness uniformity or required sample fluidity during CROF.

[0291] (vi) Shim height. In some embodiments, all shims have the same predetermined height. In some embodiments, the shims have different predetermined heights. In some embodiments, the shims can be divided into groups or regions, wherein each group or region has its own shim height. And in some embodiments, the predetermined height of the shims is the average height of the shims. In some embodiments, the shims have approximately the same height. In some embodiments, a certain number of shims have the same height.

[0292] The height of the shim is selected by the desired specified final sample thickness and the residual sample thickness. The shim height (predetermined shim height) and / or sample thickness is 3 nm or less, 10 nm or less, 50 nm or less, 100 nm or less, 200 nm or less, 500 nm or less, 800 nm or less, 1000 nm or less, 1 μm or less, 2 μm or less, 3 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, 30 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 mm or less, 2 mm or less, 4 mm or less, or a range between any two values.

[0293] The spacer height and / or sample thickness is in a preferred embodiment from 1 nm to 100 nm, in another preferred embodiment from 100 nm to 500 nm, in a separate preferred embodiment from 500 nm to 1000 nm, in another preferred embodiment from 1 μm (e.g., 1000 nm) to 2 μm, in another preferred embodiment from 2 μm to 3 μm, in another preferred embodiment from 3 μm to 5 μm, in another preferred embodiment from 5 μm to 10 μm, in another preferred embodiment from 10 μm to 50 μm, in a separate preferred embodiment from 50 μm to 100 μm.

[0294] In certain embodiments, the spacer height and / or sample thickness is (i) equal to or slightly larger than the smallest dimension of the analyte, or (ii) equal to or slightly larger than the largest dimension of the analyte. "Slightly larger" means it is approximately 1% to 5% larger, and any number in between.

[0295] In certain embodiments, the spacer height and / or sample thickness is greater than the smallest dimension of the analyte (eg, the analyte has an anisotropic shape), but less than the largest dimension of the analyte.

[0296] For example, a red blood cell has a disc shape with a minimum dimension of 2 μm (disc thickness) and a maximum dimension of 11 μm (disc diameter). In one embodiment of the present invention, the spacer is selected so that the inner surface spacing of the plate in the relevant area is 2 μm (equal to the minimum dimension) in one embodiment, 2.2 μm in another embodiment, or 3 (50). In other embodiments, the dimension is larger than the maximum dimension of the minimum dimension, but smaller than the maximum dimension of the red blood cell. Such an embodiment has certain advantages in blood cell counting. In one embodiment, for red blood cell counting, by making the inner surface spacing 2 or 3 μm, and making any number between the two values, the undiluted whole blood sample is evenly limited to the spacing of each red blood cell (RBC) without overlapping with other cells, so that the red blood cells can be accurately counted visually. (Too much overlap between RBCs can cause serious errors in counting).

[0297] In certain embodiments of the invention, the plates and spacers are used to adjust not only the thickness of the sample, but also the orientation and / or surface density of the analytes / entities in the sample when the plate is in a closed configuration. When the plate is in a closed configuration, the thinner the thickness of the sample, the less analytes / entities per unit surface area (e.g., lower surface concentration).

[0298] (VII) Transverse dimensions of the gasket. For an open gasket, its transverse dimensions can be characterized by the transverse dimensions in two orthogonal directions, x and y (sometimes referred to as width). The transverse dimensions of the gasket in each direction may be the same or different. In certain embodiments, the transverse dimension in each direction (x or y) is…

[0299] In some embodiments, the ratio of the lateral dimensions in the x- and y-directions is 1, 1.5, 2, 5, 10, 100, 500, 1000, 10,000, or a range between any two of these values. In some embodiments, different ratios are used to adjust the flow direction of the sample; for example, the larger the ratio, the flow is in one direction (the direction of the larger dimension).

[0300] In certain embodiments, the different lateral dimensions of the spacers in the x and y directions are used to (a) use the spacers as scale marks to indicate the orientation of the plate, (b) use the spacers to produce more preferred direction sample flow, or both.

[0301] In a preferred embodiment, period, width and height.

[0302] In some embodiments, all the spacers have the same shape and size. In some embodiments, each of the spacers has a different transverse size.

[0303] For a closed gasket, in some embodiments, the internal lateral shape and dimensions are selected based on the total volume of the sample to be enclosed by the closed gasket, where the volume dimensions have been described in the present invention; in some embodiments, the external shape and dimensions are selected based on the required strength to support the pressure of the liquid against the gasket and the compression pressure of the pressure plate.

[0304] (vii) Aspect ratio of height to average lateral dimension of pillar spacers. In certain embodiments, the aspect ratio of height to average lateral dimension of the columnar spacers is 100,000, 10,000, 1,000, 100, 10, 1, 0.1, 0.01, 0.001, 0.0001, 0.0001, or a range between any two.

[0305] (ix) Shim height accuracy. The height of the shims shall be precisely controlled. The relative accuracy of the shims (e.g., the ratio of the deviation to the desired shim height) is 0.001% or less, 0.01% or less, 0.1% or less, 0.5% or less, 1% or less, 2% or less, 5% or less, 8% or less, 10% or less, 15% or less, 20% or less, 30% or less, 40% or less, 50% or less, 60% or less, 70% or less, 80% or less, 90% or less, 99.9% or less, or any range thereof.

[0306] (X) Spacer spacing. The spacer can be a single spacer or multiple spacers in the relevant area of ​​the plate or sample. In some embodiments, the spacers on the plate are configured and / or arranged in an array, and the array is periodic, aperiodic, or periodic in some locations of the plate and aperiodic in other locations.

[0307] In certain embodiments, the periodic array of shims has a lattice of squares, rectangles, triangles, hexagons, polygons, or any combination thereof, where combination means that different locations of the plate have different lattices of shims.

[0308] In some embodiments, the inter-shim distance of the shim array is periodic (eg, uniform inter-shim distance) in at least one direction of the array. In some embodiments, the shim distance is configured to improve uniformity between plate spacings in the closed configuration.

[0309] The distance between adjacent gaskets (e.g., the distance between gaskets) is 1 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, 30 μm or less, 40 μm or less, 50 μm or less, 60 μm or less, 70 μm or less, 80 μm or less, 90 μm or less, 100 μm or less, 200 μm or less, 300 μm or less, 400 μm or less, or between two values.

[0310] In some embodiments, the spacer distance is 400 or less, 500 or less, 1 mm or less, 2 mm or less, 3 mm or less, 5 mm or less, 7 mm or less, 10 mm or less, or any range between values. In some embodiments, the distance between spacers is 10 mm or less, 20 mm or less, 30 mm or less, 50 mm or less, 70 mm or less, 100 mm or less, or any range between values.

[0311] The distance between adjacent gaskets (e.g., the distance between gaskets) is selected so that for given properties of the plate and the sample, in certain embodiments, in the closed configuration of the plate, the thickness variation of the sample between two adjacent gaskets is a maximum of 0.5%, 1%, 5%, 10%, 20%, 30%, 50%, 80%, or any range between the two values; or in certain embodiments, a maximum of 80%, 100%, 200%, 400%, or a range between any two values.

[0312] Obviously, in order to maintain a given sample thickness variation between two adjacent shims, a closer shim spacing is required when using a more flexible plate.

[0313] Specify the precision of the interval distance

[0314] In a preferred embodiment, the spacers are a periodic square array, wherein the spacers are pillars having a height of 2 to 4 μm, an average lateral dimension of 5 to 20 μm, and a spacer spacing of 1 μm to 100 μm.

[0315] In a preferred embodiment, the spacers are a periodic square array, wherein the spacers are pillars having a height of 2 to 4 μm, an average lateral dimension of 5 to 20 μm, and a spacer spacing of 100 μm to 250 μm.

[0316] In a preferred embodiment, the spacers are a periodic square array, wherein the spacers are pillars having a height of 4 to 50 μm, an average lateral dimension of 5 to 20 μm, and a spacer spacing of 1 μm to 100 μm.

[0317] In a preferred embodiment, the spacers are a periodic square array, wherein the spacers are pillars having a height of 4 to 50 μm, an average lateral dimension of 5 to 20 μm, and a spacer spacing of 100 μm to 250 μm.

[0318] The period of the spacer array is 1 nm to 100 nm in one preferred embodiment, 100 nm to 500 nm in another preferred embodiment, 500 nm to 1000 nm in a separate preferred embodiment, 1 μm (e.g., 1000 nm) to 2 μm in another preferred embodiment, 2 μm to 3 μm in a separate preferred embodiment, 3 μm to 5 μm in another preferred embodiment, 5 μm to 10 μm in a separate preferred embodiment, 10 μm to 50 μm in another preferred embodiment, 50 μm to 100 μm to 175 μm in a separate preferred embodiment, and 175 μm to 300 μm in a separate preferred embodiment.

[0319] (xi) Spacer density. Spacers are measured in μm 2 Greater than 1, every 10μm 2 Greater than 1, every 100 μm 2 Greater than 1, every 500μm 2 Greater than 1, every 1000μm 2 Greater than 1, every 5000μm 2 A surface density distribution of greater than 1, greater than 1 per 0.01 square millimeter, greater than 1 per 0.1 square millimeter, greater than 1 per 1 square millimeter, greater than 1 per 5 square millimeters, greater than 1 per 10 square millimeters, greater than 1 per 100 square millimeters, greater than 1 per 1000 square millimeters, greater than 1 per 10000 square millimeters, or a range between two values.

[0320] (3) The spacer is configured so as not to occupy a significant surface area (volume) within a given sample area (volume);

[0321] (xii) Spacer volume to sample volume ratio. In many embodiments, the ratio of the spacer volume (e.g., the volume of the spacer) to the sample volume (e.g., the volume of the sample), and / or the ratio of the volume of the spacer within the relative volume of the container. The relative volume of the sample to the sample is controlled to obtain certain advantages. Advantages include, but are not limited to, uniformity of sample thickness control, uniformity of analytes, sample flow characteristics (e.g., flow rate, flow direction, etc.).

[0322] In some embodiments, the ratio of the gasket volume (r) to the sample volume, and / or the ratio of the volume of the gasket within the relevant volume of the sample to the relevant volume of the sample is less than 100%, at most 99%, at most 70%, at most 50%, at most 30%, at most 10%, at most 5%, at most 3% at most 1%, at most 0.1%, at most 0.01%, at most 0.001%, or a range between two values.

[0323] (xiii) Spacers fixed to the panels. The spacing distance between the spacers and the orientation of the spacers, which are critical in the present invention, are preferably maintained during the process of moving the panels from the open configuration to the closed configuration, and / or are preferably predetermined before changing the panels from the open configuration to the closed configuration.

[0324] In certain embodiments of the present description, a shim is secured to one of the panels before the panels are brought to a closed configuration. The term "shim is secured to its respective panel" means that the shim is attached to the panel and remains attached during use of the panel. An example of "shim is secured to its respective panel" is that the shim is integrally made of one material of the panel and the position of the shim relative to the panel surface does not change. An example of "shim is not secured to its respective panel" is that the shim is bonded to the panel by an adhesive, but when the panel is in use, the adhesive is unable to hold the shim in its original position on the panel surface (e.g., the shim moves from its original position on the panel surface).

[0325] In some embodiments, at least one pad is fixed to its respective plate. In some embodiments, two pads are fixed to their respective plates. In some embodiments, most pads are fixed with their respective plates. In some embodiments, all pads are fixed with their respective plates.

[0326] In certain embodiments, the spacer is integrally secured to the plate.

[0327] In certain embodiments, the shims are secured to their respective plates by one or any combination of the following methods and / or configurations: attaching, bonding, fusing, stamping, and etching.

[0328] The term "embossing" refers to the integral fastening of the shim and the plate by impressing (eg, embossing) a piece of material to form the shim on the surface of the plate. The material may be a single layer of material or multiple layers of material.

[0329] The term "etched" means that the pad and board are integrally secured by etching a piece of material to form the pad on the board surface. The material can be a single layer of material or multiple layers of material.

[0330] The term "fused to" means that the gasket and the plate are integrally fixed by bonding them together, original materials for the gasket and the plate are fused to each other, and there is a clear material boundary fusion between the two materials afterwards.

[0331] The term "bonded to" means that the gasket and the plate are integrally fixed by bonding the gasket and the plate through adhesion.

[0332] The term "attached to" means that the gasket and the plate are connected together.

[0333] In some embodiments, the spacer and the plate are made of the same material. In other embodiments, the spacer and the plate are made of different materials. In another embodiment, the spacer and the plate are formed as one piece. In another embodiment, one end of the spacer is fixed to its corresponding plate, while the end is open to accommodate different configurations of the two plates.

[0334] In other embodiments, each shim is independently at least one of attached to, bonded to, fused to, stamped into, and etched into the respective plate. The term "independently" means that one shim is fixed to its respective plate by the same or different methods selected from the group consisting of attached to, bonded to, fused to, stamped into, and etched into the respective plate.

[0335] In some embodiments, the distance between at least two shims is predetermined ("predetermined distance between shims" means that the distance is known when a user uses the board).

[0336] In certain embodiments of all of the methods and apparatus described herein, in addition to the fixed shims, additional shims are present.

[0337] (xiv) Specific sample thickness. In the present invention, it is observed that a larger plate holding force (e.g., the force holding two plates together) can be achieved by using a smaller plate spacing (for a given sample area) or a larger sample area (for a given sample), or both.

[0338] In certain embodiments, at least one plate is transparent in an area surrounding the associated area, each plate has an inner surface configured to contact the sample in a closed structure; the inner surfaces of the plates are substantially parallel to each other in the closed state; the inner surfaces of the plates are substantially planar, except for locations with gaskets; or any combination thereof.

[0339] , the gasket can be attached to the board in various ways using photolithography, etching, imprinting (nanoimprinting), deposition, peeling, fusion or a combination thereof. In some embodiments, the gasket is directly imprinted or embossed on the board. In some embodiments, the gasket is imprinted into a material (such as plastic) deposited on the board. In some embodiments, the gasket is made by directly imprinting the surface of the CROF board. Nanoimprinting can be done using a roll-to-roll technique using a roller imprinter, or it can be rolled into a flat nanoimprint. This method has great economic advantages, thereby reducing costs.

[0340] In some embodiments, the gasket is attached to the board. The attachment can be evaporation, sticking or peeling. In the sticking process, the gasket is first made on a carrier and then the gasket is transferred from the carrier to the board. In the lift-off, a removable material is first deposited on the board and then a hole is created in the material; the bottom of the hole exposes the board surface, and then an isolation material is deposited into the hole, and then the removable material is removed, leaving only the gasket on the board surface. In some embodiments, the gasket deposited on the board is fused to the board. In some embodiments, the gasket and the board are made in a single process. The single process includes imprinting (e.g., embossing, molding) or compositing.

[0341] In some embodiments, at least two gaskets are fixed to the corresponding plates by different manufacturing methods, and the optional different manufacturing methods include at least one of deposition, bonding, fusion, imprinting and etching.

[0342] In certain embodiments, one or more spacers are secured to the respective plates by a manufacturing method of bonding, fusing, stamping or etching or any combination thereof.

[0343] In certain embodiments, the manufacturing method for forming such an integral gasket on the board includes bonding, fusing, stamping or etching methods, or any combination thereof.

[0344] B) Adapter

[0345] Details of the adapter are described in detail in various publications, including International Application No. PCT / US2018 / 017504 (Essenlix File No. ESXPCT18F04), which are hereby incorporated by reference for all purposes.

[0346] The invention described herein solves this problem by providing a system including an optical adapter and a smartphone. The optical adapter device can be mounted on a smartphone and transform it into a microscope, which can take fluorescence and brightfield images of a sample. The system can be operated conveniently and reliably by an average person anywhere. The optical adapter utilizes the existing resources of a smartphone, including a camera, light source, processor, and display, thereby providing a low-cost solution that enables users to perform brightfield and fluorescence microscopy.

[0347] In the present invention, the optical adapter device includes a holder frame mounted on the upper part of a smartphone and an optical box attached to the holder having a sample container slot and an illumination optical device. In some references (U.S. Patent No. 2016 / 029091 and U.S. Patent No. 2011 / 0292198), the optical adapter design is a whole, including both clip-on mechanical parts that can be mounted on a smartphone and functional optical elements. The problem with this design is that they need to redesign the entire optical adapter for each specific model of smartphone. However, in the present invention, the optical adapter is divided into a bracket frame that is only used to mount the smartphone and a universal optical box that contains all functional components. For smartphones of different sizes, as long as the relative positions of the camera and the light source are the same, only the bracket frame needs to be redesigned, which can save a lot of design and manufacturing costs.

[0348] The optical box of the optical adapter includes: a receiving slot that receives the sample and places it in a sample slide within the field of view and focal length of the smartphone camera; and a brightfield illumination optics for capturing a brightfield microscopic image of the sample; a fluorescence illumination optics for capturing a fluorescence microscopic image of the sample; and a lever that switches between the brightfield illumination optics and the fluorescence illumination optics by sliding inward and outward in the optical box.

[0349] The socket slot has a rubber door that completely covers the slot to prevent ambient light from entering the optical box to be collected by the camera. In U.S. Patent No. 2016 / 0290916, the sample slot is always exposed to ambient light, which does not cause much problem because it only brightens the field microscope. But the present invention can take advantage of this rubber door when performing fluorescence microscopy, because ambient light can bring a lot of noise to the camera's image sensor.

[0350] To capture good fluorescence microscopy images, it is ideal that almost no excitation light enters the camera and the camera collects only the fluorescence emitted by the sample. However, for all common smartphones, due to the large divergence angle of the light beam emitted by the light source and infrared light source, the filter placed in front of the camera cannot well block the undesired wavelength range emitted by the smartphone light source. Optical filters do not work well for uncollimated light beams. Collimating optics can be designed to collimate the light beam emitted by the smartphone light source to solve this problem, but this approach increases the size and cost of the adapter. In contrast, in the present invention, the fluorescence illumination optics enable the excitation light to illuminate the sample at a large oblique incidence angle, partially from the waveguide inside the sample slide and partially from the back side of the sample side, so that the excitation light is almost not collected by the camera to reduce the noise signal entering the camera.

[0351] Brightfield illumination optics in the adapter receive and redirect the beam from the light source so that the sample is back-illuminated at normal incidence.

[0352] Typically, the optical box also includes a lens mounted therein that aligns with the smartphone's camera and can magnify the image captured by the camera. The image captured by the camera can be further processed by the smartphone's processor and the analysis results can be output on the smartphone's screen.

[0353] In order to realize bright field illumination and fluorescence illumination simultaneously in the same optical adapter, in the present invention, a slidable lever is used. The optical element of the fluorescence illumination optical element is mounted on the lever, and when the lever is fully slid into the optical box, the fluorescence illumination optical element blocks the light path of the bright field illumination optical element and switches the illumination optical element to the fluorescence illumination optical element. And when the lever is slid out, the fluorescence illumination optical element mounted on the lever moves out of the light path and switches the illumination optical system to the bright field illumination optical system. This lever design enables the optical adapter to work in both bright field and fluorescence illumination modes without designing two different single-mode optical boxes.

[0354] The lever comprises two planes at different levels at different heights.

[0355] In some embodiments, the two planes can be connected together with a vertical rod and moved in or out of the optical box together. In some embodiments, the two planes can be separated and each plane can be moved in or out of the optical box separately.

[0356] The upper lever plane includes at least one optical element, which may be, but is not limited to, an optical filter. The upper lever plane moves below the light source, and a preferred distance between the upper lever plane and the light source is in the range of 0 to 5 mm.

[0357] A portion of the bottom lever plane is non-parallel to the image plane. The surface of the non-parallel portion of the bottom lever plane has a mirror finish with a reflectivity greater than 95%. The non-parallel portion of the bottom lever plane moves under the light source and deflects light emitted from the light source to back-illuminate the sample area directly under the camera. The preferred tilt angle of the non-parallel portion of the bottom lever plane is in the range of 45 degrees to 65 degrees, and the tilt angle is defined as the angle between the non-parallel bottom plane and the vertical plane.

[0358] A portion of the bottom lever plane is parallel to the image plane, located below the sample and 1mm to 10mm away from the sample. The surface of the parallel portion of the bottom lever plane has a high light absorption rate, and the light absorption rate is greater than 95%. This absorptive surface is to eliminate reflected light shining on the sample at a small incident angle.

[0359] In order to slide in and out to use the lever to switch the illumination optics, a plug design including a spherical plunger and a groove on the lever is used so that the lever stops at a predetermined position when it is pulled out of the adapter. This allows the user to pull the lever with any force but stops the lever in a fixed position, in which the working mode of the optical adapter is switched to bright illumination.

[0360] The sample slider fits into the socket slot to accommodate the QMAX device and place the sample within the field of view and focus of the smartphone camera in the QMAX device.

[0361] The sample slide consists of a fixed track frame and a movable arm:

[0362] The frame rail is fixedly mounted in the slot of the optical box. The rail frame has a sliding rail slot that fits the width and thickness of the QMAX device, so the QMAX device can slide along the rail. The width and height of the rail slot are carefully configured to allow the QMAX device to deviate less than 0.5mm in the direction perpendicular to the sliding direction in the sliding plane and less than 0.2mm in the thickness direction of the QMAX device.

[0363] The frame track features an open window under the field of view of the smartphone camera to allow light to illuminate the sample backwards.

[0364] The movable arm is pre-built into the sliding track slot of the track frame and moves with the QMAX device to guide the movement of the QMAX device in the track frame.

[0365] The movable arm is equipped with a stop mechanism with two predetermined stop positions. For one position, the arm will stop the QMAX device at a position where the fixed sample area on the QMAX device is directly under the smartphone camera. For the other position, the arm will stop the QMAX device at a position where the sample area on the QMAX device is not within the field of view of the smartphone and the QMAX device can be easily removed from the track slot.

[0366] The movable arm switches between the two stop positions by pressing the QMAX device and the movable arm together to the end of the track groove and then releasing it.

[0367] The movable arm can indicate whether the QMAX device is inserted in the correct orientation. One corner of the QMAX device is shaped differently than the other three right angles. The shape of the movable arm matches the shape of the specially shaped corner, so the QMAX device can only slide into the correct position in the track slot in the correct orientation.

[0368] C) Smartphone / Detection System

[0369] Details of the smartphone / detection system are described in detail in various publications, including International Application (IA) PCT / US2016 / 046437 filed on August 10, 2016, IA PCT / US2016 / 051775 filed on September 14, 2016, U.S. Provisional Application No. 62 / 456065 filed on February 7, 2017, U.S. Provisional Application Nos. 62 / 456287 and 62 / 456590 filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017, U.S. Provisional Application No. 62 / 459,544 filed on February 15, 2017, and U.S. Provisional Application Nos. 62 / 460075 and 62 / 459920 filed on February 16, 2017, respectively, which are incorporated herein by reference in their entirety for all purposes.

[0370] The devices / apparatus, systems and methods disclosed herein may include or use a Q card for sample detection, analysis and quantification. In certain embodiments, the Q card is used with an adapter that can be connected to a smartphone detection system. In certain embodiments, the smartphone includes a camera and / or an illumination source. In certain embodiments, the smartphone includes a camera that can be used to capture an image or sample when the sample is placed in the camera's field of view (e.g., via an adapter). In certain embodiments, the camera includes a set of lenses (e.g., in an iPhone TM 6). In certain embodiments, the camera includes at least two sets of lenses (e.g., in an iPhone TM 7). In certain embodiments, the smartphone includes a camera, but the camera is not used for image capture.

[0371] In some embodiments, the smartphone includes a light source such as, but not limited to, an LED (light emitting diode). In some embodiments, the light source is used to provide illumination to the sample when the sample is in the field of view of the camera (e.g., through an adapter). In some embodiments, the light from the light source is enhanced, amplified, altered, and / or optimized by optical components of the adapter.

[0372] In certain embodiments, the smartphone includes a processor configured to process information from the sample. The smartphone includes software instructions that, when executed by the processor, can enhance, amplify and / or optimize the signal (e.g., image) from the sample. The processor can include one or more hardware components, such as a central processing unit (CPU), an application specific integrated circuit (ASIC), an application specific instruction set processor (ASIP), a graphics processing unit (GPU), a physical processing unit (PPU), a digital signal processor (DSP), a field programmable gate array (FPGA), a programmable logic device (PLD), a controller, a microcontroller unit, a reduced instruction set computer (RISC), a microprocessor, etc., or any combination thereof.

[0373] In some embodiments, the smartphone includes a communication unit configured and / or used to transmit data and / or images related to the sample to another device. By way of example only, the communication unit may use a cable network, a wired network, a fiber optic network, a telecommunications network, an intranet, the Internet, a local area network (LAN), a wide area network (WAN), a wireless local area network (WLAN), a metropolitan area network (MAN), a wide area network (WAN), a public switched telephone network (PSTN), a Bluetooth network, a ZigBee network, a near field communication (NFC) network, or the like, or any combination thereof. In some embodiments, the smartphone is an iPhone TM , Android TM Phone or Windows TM Telephone.

[0374] D) Manufacturing Method The details of the manufacturing method are described in detail in various publications, including international application PCT / US2018 / 057873 filed on October 26, 2018, which is incorporated herein by reference for all purposes.

[0375] The device of the present invention can be manufactured using techniques well known in the art. The choice of manufacturing technology will depend on the material used for the device and the size of the spacer array and / or the size of the spacer. Exemplary materials for manufacturing the device of the present invention include glass, silicon, steel, nickel, polymers, such as polymethyl methacrylate (PMMA), polycarbonate, polystyrene, polyethylene, polyolefins, silicones (e.g., poly(dimethylsiloxane)), polypropylene, cis-polyisoprene (rubber), polyvinyl chloride (PVC), polyvinyl acetate (PVAc), polychloroprene (chloroprene), polytetrafluoroethylene (Teflon), polyvinylidene chloride (SaranA) and cyclic olefin polymers (COP)) and cyclic olefin copolymers (COC) and combinations thereof. Other materials are known in the art. For example, deep reactive ion etching (DRIE) is used to manufacture silicon-based devices with small gaps, small spacers, and large aspect ratios (the ratio of spacer height to lateral dimensions). Equipment for thermoforming (compression molding, injection molding) of plastics can also be used, for example, when the smallest lateral features are >20 microns and the aspect ratio of these features is ≦10.

[0376] Other methods include lithography (e.g., stereolithography or X-ray lithography), molding, imprinting, silicon micromachining, wet or dry chemical etching, milling, diamond cutting, lithography and laser shaping (LIGA) and electroplating. For example, for glass, conventional silicon manufacturing techniques of lithography followed by wet (KOH) or dry etching (etching with reactive ions and fluorine or other reactive gases) can be employed. For plastic materials with high photon absorption efficiency, techniques such as laser micromachining can be employed. Due to the continuous nature of the process, the technology is suitable for lower-volume manufacturing. For mass-produced plastic devices, thermoplastic injection molding and compression molding may be suitable. Conventional thermoplastic injection molding for large-scale manufacturing of optical discs (which retain sub-micron feature fidelity) can also be used to manufacture the devices of the present invention. For example, device features are replicated on a glass master by conventional lithography. The glass master is electroformed to produce a tough, thermal shock resistant, thermally conductive, hard mold. The mold is used as a master template for injection or compression molding the features into a plastic device. Depending on the plastic material used to manufacture the device and the requirements for optical quality and throughput of the finished product, compression molding or injection molding can be selected as the manufacturing method. Compression molding (also known as hot embossing or relief embossing) has the advantage of being compatible with high molecular weight polymers, which are very good for small structures and can replicate high aspect ratio structures, but the cycle time is longer. Injection molding is suitable for low aspect ratio structures and is best suited for low molecular weight polymers.

[0377] The device can be made in one or more parts and then assembled. The layers of the device can be bonded together by clamping, adhesives, heating, anodic bonding, or reactions between groups of surfaces (e.g., wafer bonding). Alternatively, a device with channels or gaps in more than one plane can be made as a single piece, such as using stereolithography or other three-dimensional manufacturing techniques.

[0378] In order to reduce the non-specific adsorption of cells or compounds released by lysed cells onto the surface of the device, one or more surfaces of the device can be chemically modified with non-adherent or repelling forces. The surface can be coated with a thin film coating (e.g., a monolayer) of a commercially available non-stick reagent, such as a reagent for forming a hydrogel. The chemical substances of this additional embodiment can be used to modify the surface of the device including oligoethylene glycol, fluorinated polymers, organosilanes, thiols, polyethylene glycol, hyaluronic acid, bovine serum albumin, polyvinyl alcohol, mucin, polyhydroxyethyl methacrylate, methacrylic acid PEG and agarose. Charged polymers can also be used to repel substances of opposite charge. The type of chemical substance used for repulsion and the method of attaching to the surface of the device will depend on the properties of the repelled substance and the properties of the surface and substance to which it is attached. Such surface modification techniques are well known in the art. The surface can be functionalized before or after assembling the device. The surface of the device can be coated to capture material samples, such as membrane fragments or proteins.

[0379] In certain embodiments of the present description, the method for making any Q card of the present invention may include injection molding of a first plate. In certain embodiments of the present description, the method for making any Q card of the present invention may include nanoimprinting or extrusion printing of a second plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include laser cutting of a first plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include nanoimprinting or extrusion printing of a second plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include injection molding and laser cutting of a first plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include nanoimprinting or extrusion printing of a second plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include nanoimprinting or extrusion printing to make a first plate and a second plate. In certain embodiments of the present invention, the method for making any Q card of the present invention may include: using injection molding, laser cutting of a first plate, nanoimprinting, extrusion printing, or a combination of the following to make a first plate or a second plate: its. In certain embodiments of the present invention, the method for making any Q card of the present invention may include the step of attaching hinges to the first plate and the second plate after making the first plate and the second plate.

[0380] E) Sample type and subjects

[0381] Details of the samples and subjects are described in detail in various publications, including International Application (IA) PCT / US2016 / 046437 filed on August 10, 2016, IA PCT / US2016 / 051775 filed on September 14, 2016, IA No. PCT / US201 / 017307 filed on February 7, 2018, and IA No. PCT / US2017 / 065440 filed on December 8, 2017, each of which is incorporated herein by reference for all purposes.

[0382] The sample can be obtained from a subject. The subjects described herein can be of any age and can be an adult, infant, or child. In some cases, the subject is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98 or 99 years old, or within a certain range (e.g., between 2 and 20 years old, between 20 and 40 years old or between 40 and 90 years old). A particular class of subjects that may benefit are subjects having or suspected of having an infection (e.g., a bacterial and / or viral infection).

[0383] Another special class of subjects that can benefit is a subject at a higher risk of infection. In addition, the subject treated by any method or composition described herein can be male or female. Any method, device or kit disclosed herein can also be performed on non-human subjects such as laboratory or farm animals. Non-limiting examples of non-human subjects include dogs, goats, guinea pigs, hamsters, mice, pigs, non-human primates (e.g., gorillas, apes, orangutans, lemurs, or baboons), mice, sheep, cows or zebrafish.

[0384] The devices, apparatuses, systems and methods disclosed herein can be used for samples such as, but not limited to, diagnostic samples, clinical samples, environmental samples and food samples.

[0385] For example, in certain embodiments, the devices, apparatuses, systems and methods disclosed herein are used with samples comprising cells, tissues, body fluids and / or mixtures thereof. In certain embodiments, the sample comprises a human body fluid. In certain embodiments, the sample comprises cells, tissues, body fluids, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, uterine, chyle, bells, endolymph, perilymph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, inflammatory secretions, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine and exhaled breath condensate.

[0386] In certain embodiments, the devices, apparatuses, systems and methods disclosed herein are used for environmental samples obtained from any suitable source, such as, but not limited to: rivers, lakes, ponds, oceans, glaciers, icebergs, rainwater, snow, sewage, reservoirs, tap water, drinking water, etc.; solid samples from soil, compost, sand, rocks, concrete, wood, bricks, sewage, etc.; in certain embodiments, environmental samples are freshly obtained from the source; and gaseous samples from air, underwater vents, industrial exhaust, vehicle exhaust, etc. In certain embodiments, the environmental sample is processed. For example, the sample in non-liquid form is converted into liquid form before applying the devices, apparatuses, systems and methods of the present invention.

[0387] In certain embodiments, the devices, apparatuses, systems and methods disclosed herein are used for food samples that are suitable or have the potential to become suitable for animal consumption, such as human consumption. In certain embodiments, food samples include raw materials, cooked or processed foods, animal and plant foods, pre-processed foods, and partially or completely processed foods. In certain embodiments, samples in non-liquid form are converted to liquid form before applying the subject devices, apparatuses, systems and methods.

[0388] The devices, apparatus, systems and methods can be used to analyze samples of any volume. Examples of volumes include, but are not limited to, approximately 10 mL or less, 5 mL or less, 3 mL or less, 1 microliter (uL, also referred to herein as "uL") or less, 500 uL or less, 300 uL or less, 250 uL or less, 200 uL or less, 170 uL or less, 150 uL or less, 125 uL or less, 100 uL or less, 75 uL or less, 50 uL or less, or 100 uL or less. 25uL, less than or equal to 20uL, less than or equal to 15uL, less than or equal to 10uL, less than or equal to 5uL, less than or equal to 3uL, less than or equal to 1uL, less than or equal to 0.5uL, less than or equal to 0.1uL, less than or equal to 0.05uL, less than or equal to 0.001uL, less than or equal to 0.0005uL, less than or equal to 0.0001uL, less than or equal to 10pL, less than or equal to 1pL, or a range between two values.

[0389] In certain embodiments, the volume of the sample includes, but is not limited to, approximately 100uL or less, 75uL or less, 50uL or less, 25uL or less, 20uL or less, 15uL or less, 10uL or less, 5uL or less, 3uL or less, 1uL or less, 0.5uL or less, 0.1uL or less, 0.05uL or less, 0.001uL or less, 0.0005uL or less, 0.0001uL or less, 10pL or less, 1pL or less, or a range between two of the following values. In certain embodiments, the volume of the sample includes, but is not limited to, approximately less than or equal to 10uL, less than or equal to 5uL, less than or equal to 3uL, less than or equal to 1uL, less than or equal to 0.5uL, less than or equal to 0.1uL, less than or equal to 0.05uL, less than or equal to 0.001uL, less than or equal to 0.0005uL, less than or equal to 0.0001uL, less than or equal to 10pL, less than or equal to 1pL, or a range between two of the following values.

[0390] In some embodiments, the amount of the sample is about one drop of liquid. In some embodiments, the amount of the sample is the amount collected from a punctured finger or fingertip. In some embodiments, the amount of the sample is the amount collected from a microneedle, micropipette, or intravenous aspiration.

[0391] F) Machine Learning

[0392] Details of the network are described in detail in various publications including International Application (IA) Nos. PCT / US2018 / 017504, filed February 8, 2018, and PCT / US2018 / 057877, filed October 26, 2018, which are incorporated herein by reference for all purposes.

[0393] One aspect of the present invention provides a framework for machine learning and deep learning for analyte detection and localization. A machine learning algorithm is an algorithm that can learn from data. A more rigorous definition of machine learning is "a computer program is said to learn some class of tasks T and performance measure P from experience E if its performance on tasks in T (measured by P) improves with experience E." It explores the study and construction of algorithms that can learn from data and make predictions - algorithms that make data-driven predictions or decisions by building models based on sample inputs, thereby outperforming static program instructions.

[0394] Deep learning is a special type of machine learning that is based on a set of algorithms that try to model high-level abstractions in data. In a simple case, there might be two groups of neurons: one that receives input signals and one that sends output signals. When the input layer receives input, it passes a modified version of the input to the next layer. In a deep network, there are many layers between the input and output (these layers are not made up of neurons, but they help us think that way), allowing the algorithm to use multiple layers of processing consisting of multiple linear and nonlinear transformations.

[0395] One aspect of the present invention is to provide two analyte detection and localization methods. The first method is a deep learning method, and the second method is a combination of deep learning and computer vision methods.

[0396] (i) Deep Learning Approach. In the first approach, the disclosed analyte detection and localization workflow consists of two phases: training and prediction. We describe the training and prediction phases in the following paragraphs.

[0397] (a) Training phase

[0398] During the training phase, the annotated training data is fed into a convolutional neural network. A convolutional neural network is a specialized neural network designed to process data with a grid-like, feed-forward, and hierarchical network topology. Examples of data include time series data, which can be viewed as a 1D grid sampled at fixed time intervals, and image data, which can be viewed as a 2D grid of pixels. Convolutional networks have been successful in real-world applications. The name "convolutional neural network" indicates that the network employs a mathematical operation called convolution. Convolution is a special type of linear operation. Convolutional networks are simply neural networks that use convolution instead of general matrix multiplication in at least one of their layers.

[0399] The machine learning model receives one or more sample images containing analytes acquired by the imager on a sample-mounted QMAX device as training data. The training data is annotated with the analytes to be analyzed, wherein the annotations indicate whether the analytes are in the training data and their locations in the image. The annotations can be in the form of tight bounding boxes, which completely contain the analytes or the central location of the analytes. In the latter case, the central location is further converted to a circle covering the analyte or a Gaussian kernel in the dot map.

[0400] Training machine learning models presents two challenges when the size of the training data is large: annotation (usually done by humans) is time-consuming, and training is computationally expensive. To overcome these challenges, the training data can be divided into smaller patches, and then these patches or parts of patches are annotated and trained. The term "machine learning" can refer to algorithms, systems, and devices in the field of artificial intelligence that often use statistical techniques and artificial neural networks that are trained from data without being explicitly programmed.

[0401] The annotated images are fed to the Machine Learning (ML) training module and the model trainer in the Machine Learning module will train the ML model from the training data (annotated sample images). The input data will be fed to the model trainer in multiple iterations until a specific stopping criterion is met. The output of the ML training module is the ML model - a computational model that is built by learning from data according to the training process in machine learning that enables the computer to perform certain tasks on its own (e.g., detecting and classifying objects).

[0402] The trained machine learning model is applied in the prediction (or inference) stage of the computer. Examples of machine learning models include ResNet, DenseNet, etc., which are also called "deep learning models" due to the depth of the connection layers in their network structure. In some embodiments, the Caffe library with a fully convolutional network (FCN) is used for model training and prediction, and other convolutional neural network architectures and libraries such as TensorFlow may also be used.

[0403] The training phase generates a model that will be used in the prediction phase. This model can be reused in the prediction phase to analyze inputs. Therefore, the compute unit only needs access to the generated model. It does not need access to the training data, nor does the training phase need to be run again on the compute unit.

[0404] (b) Prediction stage

[0405] In the prediction / inference phase, the detection component is applied to the input image, which is then fed into the prediction (inference) module pre-loaded with the training model generated from the training phase. The output of the prediction phase can be a bounding box containing the detected analytes and their center locations, or a dot map indicating the location of each analyte, or a heat map containing information about the detected analytes.

[0406] When the output of the prediction phase is a list of bounding boxes, the number of analytes in the sample image to be detected is characterized by the number of bounding boxes detected. When the output of the prediction phase is a spot map, the number of analytes in the sample image for analysis is characterized by the integral of the spot map. When the output of the prediction is a heat map, the localization component is used to identify the locations, and the number of detected analytes is characterized by the entries of the heat map.

[0407] One embodiment of the localization algorithm is to sort the heat map values ​​from the highest value to the lowest value into a one-dimensional ordered list. Then select the pixel with the highest value and remove the pixel and its neighboring pixels from the list. Repeat this process to select the pixel with the highest value in the list until all pixels are removed from the list.

[0408] In the detection component using heatmap, the input image is fed into the convolutional neural network along with the model generated from the training phase and the output of the detection phase is pixel-wise predictions in the form of a heatmap. The heatmap can be of the same size as the input image or a scaled-down version of the input image and it is the input to the localization component. We disclose an algorithm to localize the center of the analyte. The main idea is to iteratively detect local peaks from the heatmap. After the peak is localized, we calculate the local area around the peak, but with smaller values. We remove that area from the heatmap and find the next peak from the remaining pixels. The process is repeated until all the pixels are removed from the heatmap.

[0409] In certain embodiments, the present invention provides a localization algorithm to sort the heat map values ​​from the highest value to the lowest value into a one-dimensional ordered list. The pixel with the highest value is then selected and the pixel and its neighboring pixels are removed from the list. This process is repeated to select the pixel with the highest value in the list until all pixels are removed from the list.

[0410] Global search algorithm (heat map)

[0411]

[0412]

[0413] After sorting, the heatmap is a one-dimensional ordered list where the heatmap values ​​are ordered from highest to lowest. Each heatmap value is associated with its corresponding pixel coordinate. The first item in the heatmap is the item with the highest value, which is the output of the pop(heatmap) function. A disk is created with the center being the pixel coordinate of the disk with the highest heatmap value. All heatmap values ​​whose pixel coordinates are inside the disk are then removed from the heatmap. The algorithm repeatedly pops the maximum value in the current heatmap and removes the disks around it until all items are removed from the heatmap.

[0414] In an ordered list heatmap, each item has knowledge of the ongoing and subsequent items. When removing an item from an ordered list, we make the following changes:

[0415] Assume the removed item is x r , whose continuation term is x p , whose subsequent term is x f .

[0416] For the forward term x p , redefine its successor to be the successor of the deleted term. Therefore, x p The next term is now x f .

[0417] For deleting item x r , undefines its continuation and successor, thereby removing it from the ordered list.

[0418] For the following items x f , redefines its ongoing project as the ongoing project of the removed project. Therefore, x f The progressive term is now x p .

[0419] After all items are removed from the ordered list, the localization algorithm is complete. The number of items in the site will be the number of analytes and the location information is the pixel coordinates of each s in the site.

[0420] Another embodiment searches for local peaks, which is not the local peak with the highest heatmap value. To detect each local peak, we start from a random starting point and search for the local maximum. Once a peak is found, we calculate the local area around the peak, but with a smaller value. We remove that area from the heatmap and find the next peak from the remaining pixels. This process is repeated until all pixels are removed from the heatmap.

[0421] Local search algorithm (s, heatmap)

[0422]

[0423] This is a breadth-first search algorithm starting from s, where the conditions for visiting points are changed: if heatmap[p]>0 and heatmap[p]<=heatmap[q]. Therefore, every pixel in the cover has a non-descending path leading to the local peak s.

[0424] Algorithm localization (heat map)

[0425]

[0426] (ii) A hybrid of deep learning and computer vision approaches. In the second approach, detection and localization are achieved by computer vision algorithms, and classification is achieved by deep learning algorithms, where the computer vision algorithm detects and localizes possible candidates for the analyte, and the deep learning algorithm classifies each possible candidate as a true analyte or a false analyte. The locations of all true analytes (along with the total number of true analytes) will be recorded as output.

[0427] (a) Detection. Computer vision algorithms detect possible candidates based on the characteristics of the analyte, including but not limited to intensity, color, size, shape, distribution, etc. Preprocessing schemes can improve the detection results. Preprocessing schemes include contrast enhancement, histogram adjustment, color enhancement, noise reduction, smoothing, defocusing, etc. After preprocessing, the input image is sent to the detector. The detector informs about the possible presence of analytes and gives an estimate of their location. Detection can be based on the structure of the analyte (e.g. edge detection, line detection, circle detection, etc.), connectivity (e.g. blob detection, connected components, contour detection, etc.), intensity, color, shape using schemes such as adaptive thresholding, etc.

[0428] (b) Localization. After detection, a computer vision algorithm localizes each possible analyte candidate by providing its boundaries or a tight bounding box containing the boundaries. This can be achieved through object segmentation algorithms such as adaptive thresholding, background subtraction, flood fill, mean shift, watershed, etc. Many times, localization can be combined with detection to produce a detection result and the location of each analyte candidate.

[0429] (c) Classification. Deep learning algorithms such as convolutional neural networks achieve state-of-the-art visual classification. We employ deep learning algorithms to classify each possible analyte. Various convolutional neural networks can be used for analyte classification, such as VGGNet, ResNet, MobileNet, DenseNet, etc.

[0430] Given each possible analyte candidate, the deep learning algorithm computes layers of neurons through convolutional filters and nonlinear filters to extract high-level features that distinguish analytes from non-analytes. The layers of the full convolutional network incorporate high-level features into the classification results to distinguish whether there is a real analyte, or the possibility of the analyte's existence.

[0431] G) Applications, biological / chemical biomarkers and health conditions

[0432] Applications of the present invention include, but are not limited to (a) detection, purification and quantification of compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, mental illness, neuropsychiatric diseases and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in the environment (such as water, soil or biological samples); (c) detection and quantification of compounds or biological samples that pose a threat to food safety or national security, such as toxic waste, anthrax; (d) measurement of important parameters in chemical medicine or physiological monitors, such as glucose, blood oxygen levels, total blood cell count, (e) detection and quantification of specific DNA or RNA from biological samples (such as cells, viruses, body fluids), (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genome analysis or (g) detection of reaction products, such as during drug synthesis or purification.

[0433] Detection can be performed in various sample matrices, such as cells, tissues, body fluids and feces. Detectable body fluids include, but are not limited to, amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma, serum, etc.), breast milk, cerebrospinal fluid (CSF), cerumen (ear wax), chyle, chyme, endolymph, perilymph, feces, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, sebum (skin oil), semen, sputum, sweat, synovial fluid, tears, vomitus, urine and exhaled condensate. In certain embodiments, the sample comprises a human body fluid. In certain embodiments, the sample comprises cells, tissue, body fluids, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, uterine, chyle, chyme, endolymph, peripheral lymph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine, and exhaled condensate.

[0434] In embodiments, the sample is at least one of a biological sample, an environmental sample, and a biochemical sample.

[0435] The devices, systems and methods of the present invention can be used in a variety of different applications in a variety of fields where it is necessary to determine the presence and / or quantification of one or more analytes in a sample. For example, the subject methods can be used to detect proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, etc. The various fields include, but are not limited to, human, veterinary, agricultural, food, environmental, drug testing, etc.

[0436] In certain embodiments, the methods of the present invention can be used to detect nucleic acids, proteins or other biomolecules in a sample. The methods can include detecting a set of biomarkers in a sample, such as two or more different protein or nucleic acid biomarkers. For example, the methods can be used to rapidly detect two or more disease biomarkers in a biological sample for clinical purposes, such as can be used to diagnose a disease condition in a subject or in the ongoing management and treatment of a sick subject. As described above, communication with a physician or other health care provider can better ensure that the physician or other health care provider is aware of the patient's condition and can take appropriate action accordingly.

[0437] Applications of the apparatus, systems and methods of the present invention using CROF devices include, but are not limited to, (a) detection, purification and quantification of compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, mental illness, neuropsychiatric diseases and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in environments such as water, soil or biological samples such as tissues, body fluids, (c) detection and quantification of compounds or biological samples that pose a threat to food safety or national security (such as toxic waste, anthrax, etc.), (d) quantification of important parameters in medical or physiological monitors, such as glucose, blood oxygen levels, total blood cell counts, (e) detection and quantification of DNA or RNA in specific biological samples (such as cells, viruses, body fluids), (f) sequencing and comparing genetic sequences of DNA in chromosomes and mitochondria for genomic analysis, or (g) detection of reaction products during, for example, drug synthesis or purification. Some specific applications of the apparatus, systems and methods of the present invention are now described in further detail.

[0438] Applications of the present invention include, but are not limited to (a) detection, purification and quantification of compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, mental illness, neuropsychiatric diseases and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in the environment (such as water, soil or biological samples); (c) detection and quantification of compounds or biological samples that pose a threat to food safety or national security, such as toxic waste, anthrax; (d) quantification of important parameters in medical or physiological monitors, such as glucose, blood oxygen levels, total blood cell count, (e) detection and quantification of specific DNA or RNA from biological samples (such as cells, viruses, body fluids), (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genome analysis or (g) detection of reaction products, such as in drug synthesis or purification processes.

[0439] Embodiments of the devices, systems and methods of the present invention may include: a) obtaining a sample, b) adding the sample to a CROF device containing a capture agent that binds to a target analyte under conditions suitable for the analyte to bind to the capture agent; c) cleaning the CROF device, and d) reading the CROF device to obtain a measurement of the amount of the analyte in the sample. In certain embodiments, the analyte can be a biomarker, an environmental marker, or a food marker. In some cases, the sample is a liquid sample and can be a diagnostic sample (e.g., saliva, serum, blood, sputum, urine, sweat, tears, semen, or mucus); an environmental sample obtained from a river, ocean, lake, rainwater, snow, sewage, sewage treatment runoff, agricultural runoff, industrial runoff, tap water, or drinking water; or a food sample obtained from tap water, drinking water, prepared food, processed food, or raw food.

[0440] In any embodiment, the CROF device may be placed in a microfluidic device, and step b) may include adding the sample to the microfluidic device including the CROF device.

[0441] In any embodiment, step d) may include detecting a fluorescent or luminescent signal from the CROF device.

[0442] In any embodiment, step d) may include reading the CROF device using a handheld device configured to read the CROF device. The handheld device may be a mobile phone, such as a smartphone.

[0443] In any embodiment, the CROF device can include a labeling agent that can bind to the analyte-capture agent complex on the CROF device.

[0444] In any embodiment, the apparatus, system and method of the present invention may further comprise the step of adding a labeling agent bound to the analyte-capture agent complex on the CROF device to the CROF device between steps c) and d) and then washing the CROF device.

[0445] In any embodiment, the reading step d) may include reading an identifier of the CROF device. The identifier may be an optical barcode, a radio frequency ID tag, or a combination thereof.

[0446] In any embodiment, the devices, systems and methods of the present invention may further include adding a control sample to a CROF control device comprising a capture agent that binds to the analyte, wherein the control sample includes a known detectable amount of the analyte, and reading the control CROF device to obtain a control measurement value of the known detectable amount of the analyte in the sample.

[0447] In any embodiment, the sample can be a diagnostic sample obtained from a subject, the analyte can be a biomarker, and the measured amount of the analyte in the sample can be used in the diagnosis of a disease or condition.

[0448] In any embodiment, the devices, systems and methods of the present invention may further include receiving or providing a report to the subject indicating the measured amount of the biomarker and the range of measured values ​​of the biomarker in individuals with no or low risk of the disease or condition, wherein the comparison of the measured amount of the biomarker to the range of measured values ​​can be used for diagnosis of the disease or condition.

[0449] In any embodiment, the devices, systems and methods of the present invention may also include diagnosing the subject based on information including the measured amount of the biomarker in the sample. In some cases, the diagnosing step includes sending data including the measured amount of the biomarker to a remote location and receiving a diagnosis based on the information including the measured value from the remote location.

[0450] In any embodiment, step b) can include isolating miRNA from the sample to produce an isolated miRNA sample, and adding the isolated miRNA sample to an array of disk-coupled dots-on-pillar antennas (CROF device).

[0451] In any embodiment, the method may include receiving or providing a report indicating the safety or harmfulness of the subject to be exposed to the environment from which the sample was obtained.

[0452] In any embodiment, the method may include: sending data including the measured amount of the environmental marker to a remote location; and receiving a report indicating the safety or harmfulness of the subject to be exposed to the environment from which the sample was obtained.

[0453] In any embodiment, the CROF device array can include a plurality of capture agents that each bind to an environmental marker, and wherein step d) can include obtaining a measure of the amount of the plurality of environmental markers in the sample.

[0454] In any embodiment, the sample can be a food sample, wherein the analyte can be a food marker, and wherein the amount of the food marker in the sample can be correlated to the safety of the food for consumption.

[0455] In any embodiment, the method may include receiving or providing a report indicating the safety or harmfulness of consumption by the subject of the food from which the sample was obtained.

[0456] In any embodiment, the method may include transmitting data including the measured amounts of food markers to a remote location and receiving a report indicating the safety or harmfulness of the subject consuming the food from which the sample was obtained.

[0457] In any embodiment, the CROF device array may include a plurality of capture agents each bound to a food marker, wherein said obtaining may include obtaining a measure of the amount of the plurality of food markers in the sample, and wherein the plurality of food markers in the sample may be correlated to the safety of the edible food.

[0458] Also provided herein are kits useful in practicing the devices, systems and methods of the invention.

[0459] The sample size may be about one drop of sample. The sample size may be the amount collected from a punctured finger or fingertip. The sample size may be the amount collected from a microneedle or intravenous aspiration.

[0460] Once a sample is obtained from the source, it can be used without further processing, or it can undergo processing such as enrichment of target analytes, removal of large particulate matter, dissolution or resuspending of solid samples, etc.

[0461] Any suitable method of adding the sample to the CROF device may be used. Suitable methods may include using a pipette, dropper, syringe, etc. In certain embodiments, the CROF device may be placed on a support to form a drop rod configuration, and as described below, the sample may be added to the CROF device by immersing the sample receiving area on the drop rod into the sample.

[0462] Samples may be collected once or multiple times. Samples collected over time may be aggregated and / or processed separately (by applying to a CROF device and obtaining measurements of the amount of analyte in the sample, as described herein). In some cases, measurements obtained over time may be aggregated and used for longitudinal analysis over time to facilitate screening, diagnosis, treatment, and / or disease prevention.

[0463] As described above, the CROF device may be washed in any convenient manner to remove unbound sample components. In certain embodiments, the surface of the CROF device is washed with a binding buffer to remove unbound sample components.

[0464] The detectable labeling of the analyte can be accomplished by any convenient method. The analyte can be labeled directly or indirectly. In direct labeling, the analyte in the sample is labeled before the sample is added to the CROF device. In indirect labeling, as described below, the unlabeled analyte in the sample is labeled after the sample is added to the CROF device to capture the unlabeled analyte.

[0465] Samples from subjects, health conditions of subjects, and other applications of the invention are further described below. Exemplary samples, health conditions, and applications are also disclosed in, for example, U.S. Publication Nos. 2014 / 0154668 and 2014 / 0045209, which are incorporated herein by reference.

[0466] The present invention can be used in applications for a variety of purposes, among which such applications are generally analyte detection applications, where the presence of a specific analyte in a given sample is detected at least qualitatively, if not quantitatively. Protocols for performing analyte detection assays are well known to those skilled in the art and need not be described in detail herein. Typically, a sample that may contain an analyte is contacted with the surface of a target nanosensor under conditions sufficient to bind the analyte to its respective capture agent bound to the sensor. The capture agent has a highly specific affinity for the target targeting molecule. This affinity can be an antigen-antibody reaction in which an antibody binds to a specific epitope on an antigen, or a DNA / RNA or DNA / RNA hybridization reaction with sequence specificity between two or more complementary chains. Therefore, if the target analyte is present in the sample, it is likely to bind to the sensor at the location of the capture agent and form a complex on the sensor surface. That is, the captured analyte is fixed to the sensor surface. After removing the unbound analyte, the presence of the bound complex on the sensor surface (e.g., the fixed target analyte) is then detected, for example, using a labeled second capture agent.

[0467] Applications for the detection of specific analytes to be detected include hybridization assays using nucleic acid capture agents and protein binding assays using polypeptides, such as antibodies. In these assays, the sample is first prepared, and then after the sample is prepared, the sample is contacted with the nanosensor of the present invention under specific binding conditions, thereby forming a complex between the target nucleic acid or polypeptide (or other molecule) complementary to the capture agent attached to the sensor surface.

[0468] In one embodiment, the capture oligonucleotide is a synthetic single-stranded DNA of 20-100 bases in length, one end of which is thiolated. These molecules are fixed to the surface of the nanodevice to capture the target single-stranded DNA (on which there may be a sequence complementary to the immobilized capture DNA of at least 50bp in length). After the hybridization reaction, a detection single-stranded DNA (20-100bp in length) is added, whose sequence is complementary to the unoccupied nucleic acid of the target DNA, thereby hybridizing with the target. One end of the detection DNA is coupled to a fluorescent marker, and the emission wavelength of the fluorescent marker is within the plasma resonance range of the nanodevice. Therefore, by detecting the fluorescent emission emitted from the surface of the nanodevice, the targeted single-stranded DNA can be accurately detected and quantified. The length of the capture and detection DNA determines the melting temperature (the nucleotide chains will separate above the above melting temperature) and the degree of mismatch (the more common the chain, the lower the mismatch rate).

[0469] One consideration in selecting the length of complementary binding depends on minimizing the mismatch rate while keeping the melting temperature as high as possible. In addition, the total length of the hybridization length is determined to achieve optimal signal amplification.

[0470] The sensor of the present invention can be used in a method for diagnosing a disease or condition, comprising: (a) obtaining a liquid sample from a patient suspected of having a disease or condition, (b) contacting the sample with a nanosensor of the present invention, wherein a capture reagent on the nanosensor specifically binds to a biomarker of the disease, and capture occurs under conditions suitable for the biomarker to specifically bind to the capture reagent; (c) removing any biomarker that is not bound to the capture reagent; (d) reading an optical signal from the biomarker that is still bound to the nanosensor, wherein the optical signal indicates that the patient has a disease or condition, wherein the method further comprises labeling the biomarker with a luminescent marker before or after binding to the capture reagent. As will be described in more detail below, the patient may be suspected of having cancer, and the antibody binds to the cancer biomarker. In other embodiments, the patient is suspected of having a neurological disease, and the antibody binds to a biomarker of a neurological disease.

[0471] Applications of the sensors of the present invention include, but are not limited to (a) detection, purification and quantification of chemical compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, mental illness, neuropsychiatric diseases and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in the environment (such as water, soil or biological samples); (c) detection and quantification of compounds or biological samples that pose a threat to food safety or national security, such as toxic waste, anthrax; (d) quantification of important parameters in medical or physiological monitors, such as glucose, blood oxygen levels, total blood cell counts, (e) detection and quantification of specific DNA or RNA in biological samples (such as cells, viruses, body fluids), (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genome analysis, or (g) detection of reaction products, such as during drug synthesis or purification.

[0472] Detection can be performed in a variety of sample matrices, such as cells, tissues, body fluids and feces. Body fluids of interest include, but are not limited to, amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma, serum, etc.), breast milk, cerebrospinal fluid (CSF), cerumen (ear wax), chyle, chyme, endolymph, perilymph, feces, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, sebum (skin oil), semen, sputum, sweat, synovial fluid, tears, vomitus, urine and exhaled condensate.

[0473] In certain embodiments, the biosensor of the present invention can be used to diagnose pathogen infection by detecting a target nucleic acid of the pathogen in a sample. The target nucleic acid can be, for example, selected from a virus selected from human immunodeficiency virus 1 and 2 (HIV-1 and HIV-2), human T-cell leukemia virus and 2 (HTLV-1 and HTLV-2), respiratory syncytial virus (RSV), adenovirus, hepatitis B virus (HBV), hepatitis C virus (HCV), Epstein-Barr virus (EBV), human papillomavirus (HPV), varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex virus 1 and 2 (HSV-1 and HSV-2), human herpes virus 8 (HHV-8, also known as Kaposi's sarcoma herpesvirus) and flaviviruses, including yellow fever virus, dengue virus, Japanese encephalitis virus, West Nile virus and Ebola virus. However, the invention is not limited to the detection of nucleic acids, for example DNA or RNA sequences, from the aforementioned viruses, but can be applied without problems to other pathogens of importance for veterinary and / or human medicine.

[0474] Human papillomavirus (HPV) is further subdivided into more than 70 different types based on their DNA sequence homology. These types cause different diseases. HPV types 1, 2, 3, 4, 7, 10 and 26-29 cause benign warts. HPV types 5, 8, 9, 12, 14, 15, 17 and 19-25 and 46-50 cause damage in patients with weakened immune systems. Types 6, 11, 34, 39, 41-44 and 51-55 cause benign condyloma acuminatum on the mucous membranes of the genitals and respiratory tract. HPV types 16 and 18 are of special medical importance because they cause epithelial dysplasia of the genital mucosa and account for a large proportion of invasive carcinomas of the cervix, vagina, vulva and anal canal. DNA integration of human papillomavirus is considered to play a decisive role in the carcinogenicity of cervical cancer. Human papillomavirus can be detected from the DNA sequences of, for example, the capsid proteins L1 and L2. Therefore, the method of the present invention is particularly suitable for detecting DNA sequences of HPV types 16 and / or 18 in tissue samples to assess the risk of cancer occurrence.

[0475] In some cases, nanosensors can be used to detect biomarkers present at low concentrations. For example, nanosensors can be used to detect cancer antigens in readily accessible body fluids (e.g., blood, saliva, urine, tears, etc.), to detect biomarkers of tissue-specific diseases in readily accessible body fluids (e.g., biomarkers for neurological diseases (e.g., Alzheimer's disease antigens), to detect infections (especially detection of low titer latent viruses such as HIV), to detect fetal antigens in maternal blood, and to detect exogenous compounds (e.g., drugs or pollutants).

[0476] The following table provides a list of protein biomarkers and their associated diseases that can be detected using the nanosensors of the present invention (when used in conjunction with appropriate monoclonal antibodies). A potential source of the biomarker is also indicated in the table (e.g., "CSF"; cerebrospinal fluid). In many cases, the biosensors of the present invention can detect those biomarkers in a body fluid different from the indicated body fluid. For example, biomarkers present in CSF can be identified in urine, blood, or saliva.

[0477] H) Practicality

[0478] The method of the present invention can be used in a variety of different applications where it is necessary to determine the presence and / or quantification of one or more analytes in a sample. For example, the subject method can be used to detect proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, etc.

[0479] In certain embodiments, the methods of the present invention can be used to detect nucleic acids, proteins or other biomolecules in a sample. The methods can include detecting a set of biomarkers in a sample, such as two or more different protein or nucleic acid biomarkers. For example, the methods can be used for rapid clinical detection of two or more disease biomarkers in a biological sample, such as can be used to diagnose a disease condition in a subject or in the ongoing management or treatment of a disease condition in a subject. As described above, communicating with a physician or other health care provider can better ensure that the physician or other health care provider understands the condition and is therefore more likely to take appropriate action.

[0480] Applications of the apparatus, systems and methods of the present invention using CROF devices include, but are not limited to, (a) detection, purification and quantification of chemical compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, mental illness, neuropsychiatric diseases and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in environments such as water, soil or biological samples such as tissues, body fluids, (c) detection of compounds or biological samples that pose a threat to food safety or national security (such as toxic waste, anthrax, etc.), (d) quantification of important parameters in medical or physiological monitors, such as glucose, blood oxygen levels, total blood cell counts, (e) detection and quantification of specific DNA or RNA in biological samples such as cells, viruses, body fluids, (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genomic analysis, or (g) detection of reaction products, such as during drug synthesis or purification. Some specific applications of the apparatus, systems and methods of the present invention are now described in further detail.

[0481] I) Diagnostic methods

[0482] In certain embodiments, the methods of the present invention can be used to detect biomarkers. In certain embodiments, the devices, systems and methods of the present invention using CROF are used to detect the presence or absence of specific biomarkers, as well as the increase or decrease in the concentration of specific biomarkers in blood, plasma, serum. Or other body fluids or excretions, such as but not limited to urine, blood, serum, plasma, saliva, semen, prostatic fluid, nipple aspirate, tears, sweat, feces, cheek swabs, cerebrospinal fluid, cell lysate samples, amniotic fluid, gastrointestinal fluid, biopsy tissue, etc. Therefore, samples, such as diagnostic samples, can include various fluid or solid samples.

[0483] In some cases, the sample can be a body fluid sample from a subject to be diagnosed. In some cases, a solid or semi-solid sample can be provided. The sample can include tissues and / or cells collected from a subject. The sample can be a biological sample. Examples of biological samples can include, but are not limited to, blood, serum, plasma, nasal swabs, nasopharyngeal washes, saliva, urine, gastric juice, spinal fluid, tears, feces, mucus, sweat, earwax, oil, glandular secretions, cerebrospinal fluid, tissue, semen, vaginal fluid, tissue fluid derived from tumor tissue, eye fluid, spinal fluid, throat swabs, breath, hair, nails, skin, biopsy, placental fluid, amniotic fluid, umbilical cord blood, lymph, cavity fluid, sputum, pus, microbial flora, meconium, breast milk, exhaled condensate and / or other excreta. The sample can include nasopharyngeal washes. Nasal swabs, throat swabs, stool samples, hair, nails, earwax, breath and other solid, semi-solid or gas samples can be processed in extraction buffers, such as fixed time or variable time for their analysis. If desired, the extraction buffer or an aliquot thereof may then be processed similarly to other fluid samples.Examples of tissue samples from a subject may include, but are not limited to, connective tissue, muscle tissue, neural tissue, epithelial tissue, cartilage, cancerous samples, or bone.

[0484] In some cases, the subject from which the diagnostic sample is obtained may be a healthy individual, or may be an individual who is at least suspected of having a disease or health condition. In some cases, the subject may be a patient.

[0485] In certain embodiments, the CROF device includes a capture agent that is configured to specifically bind to a biomarker in a sample provided by a subject. In certain embodiments, the biomarker can be a protein. In certain embodiments, the biomarker protein is specifically bound by an antibody capture agent present in the CROF device. In certain embodiments, the biomarker is an antibody that specifically binds to an antigen capture agent present in the CROF device. In certain embodiments, the biomarker is a nucleic acid that specifically binds to a nucleic acid capture agent that is complementary to one or both strands of a double-stranded nucleic acid biomarker or complementary to a single-stranded biomarker. In certain embodiments, the biomarker is a nucleic acid that is specifically bound by a nucleic acid binding protein. In certain embodiments, the biomarker is specifically bound by an aptamer.

[0486] The presence or absence of a biomarker or a significant change in the concentration of a biomarker can be used to diagnose disease risk, the presence of a disease in an individual, or to adjust the treatment of a disease in an individual. For example, the presence of a specific biomarker or group of biomarkers can affect the choice of drug treatment or dosing regimen given to an individual. When evaluating potential drug therapies, biomarkers can be used as surrogates for natural endpoints (such as survival or irreversible morbidity). If a treatment changes a biomarker that is directly related to improved health status, the biomarker can be used as a surrogate endpoint to evaluate the clinical benefit of a specific treatment or dosing regimen. Therefore, personalized diagnosis and treatment based on a specific biomarker or a group of biomarkers detected in an individual is facilitated by the methods of the present invention. In addition, as described above, the high sensitivity of the devices, systems and methods of the present invention facilitates early detection of biomarkers associated with the disease. Due to the ability to detect multiple biomarkers using mobile devices (such as smartphones) combined with sensitivity, scalability and ease of use, the methods disclosed in the present invention can be used in portable and point-of-care or close-range patient molecular diagnostics.

[0487] In certain embodiments, the methods of the present invention can be used to detect biomarkers of a disease or disease state. In certain cases, the methods of the present invention can be used to detect biomarkers to characterize cell signaling pathways and intracellular communications for drug discovery and vaccine development. For example, the methods can be used to detect and / or quantify the amount of biomarkers in diseased, healthy or benign samples. In certain embodiments, the methods of the present invention can be used to detect biomarkers of infectious diseases or disease states. In certain cases, the biomarkers can be molecular biomarkers, such as but not limited to proteins, nucleic acids, carbohydrates, small molecules, etc.

[0488] The methods of the invention can be used in diagnostic assays, such as, but not limited to, the following: as described above, detection and / or quantification of biomarkers; screening tests, in which samples from asymptomatic subjects are examined periodically; prognostic assays, in which the presence and / or amount of biomarkers is used to predict the likely course of a disease; stratified analyses, in which a subject's response to different drug treatments can be predicted; efficacy analyses, in which the efficacy of a drug treatment is monitored; and the like.

[0489] In certain embodiments, the biosensor of the present invention can be used to diagnose pathogen infection by detecting a target nucleic acid of the pathogen in a sample. The target nucleic acid can be, for example, selected from a virus selected from human immunodeficiency virus 1 and 2 (HIV-1 and HIV-2), human T-cell leukemia virus and 2 (HTLV-1 and HTLV-2), respiratory syncytial virus (RSV), adenovirus, hepatitis B virus (HBV), hepatitis C virus (HCV), Epstein-Barr virus (EBV), human papillomavirus (HPV), varicella zoster virus (VZV), cytomegalovirus (CMV), herpes simplex virus 1 and 2 (HSV-1 and HSV-2), human herpes virus 8 (HHV-8, also known as Kaposi's sarcoma herpesvirus) and flaviviruses, including yellow fever virus, dengue virus, Japanese encephalitis virus, West Nile virus and Ebola virus. However, the invention is not limited to the detection of nucleic acids, for example DNA or RNA sequences, from the aforementioned viruses, but can be applied without problems to other pathogens of importance for veterinary and / or human medicine.

[0490] Human papillomavirus (HPV) is further subdivided into more than 70 different types based on their DNA sequence homology. These types cause different diseases. HPV types 1, 2, 3, 4, 7, 10 and 26-29 cause benign warts. HPV types 5, 8, 9, 12, 14, 15, 17 and 19-25 and 46-50 cause damage in patients with weakened immune systems. Types 6, 11, 34, 39, 41-44 and 51-55 cause benign condyloma acuminatum on the mucous membranes of the genitals and respiratory tract. HPV types 16 and 18 are of special medical importance because they cause epithelial dysplasia of the genital mucosa and account for a large proportion of invasive carcinomas of the cervix, vagina, vulva and anal canal. DNA integration of human papillomavirus is considered to play a decisive role in the carcinogenicity of cervical cancer. Human papillomavirus can be detected from the DNA sequences of, for example, the capsid proteins L1 and L2. Therefore, the method of the present invention is particularly suitable for detecting DNA sequences of HPV types 16 and / or 18 in tissue samples to assess the risk of cancer occurrence.

[0491] Other pathogens that can be detected in diagnostic samples using the devices, systems and methods of the present invention include, but are not limited to: varicella zoster; Staphylococcus epidermidis, Escherichia coli, methicillin-resistant Staphylococcus aureus (MSRA), Staphylococcus aureus, Staphylococcus hominis, Enterococcus faecalis, Pseudomonas aeruginosa, Tinea capitis, Staphylococcus worderi, Klebsiella pneumoniae, Haemophilus influenzae, Staphylococcus aegypti, Streptococcus pneumoniae and Candida albicans; Neisseria gonorrhoeae gorrhoeae), syphilis (Treponenapallidμm), Clamyda tracomitis, non-gonococcal urethritis (UreaplasmUrzyplasmμm), chancroid (Haemophilusducreyi), trichomoniasis (Trichomonasvaginalis); Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus (MSRA), Klebsiella pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Stenotrophomonas maltophilia, Haemophilus parahaemolyticus, Escherichia coli, Enterococcus faecalis, Enterococcus faecalis, Enterococcus, Enterococcus faecalis, Citrobacter freundii, Enterococcus, Klebsiella oxytoca, Pseudomonas, Neisseria meningitidis, Streptococcus pyogenes, Pneumocystis carinii, Legionella pneumophila, Mycoplasma pneumoniae, and Mycobacterium tuberculosis, etc.

[0492] In some cases, CROF devices can be used to detect low concentrations of biomarkers. For example, CROF devices can be used to detect cancer antigens in readily accessible body fluids (e.g., blood, saliva, urine, tears, etc.), to detect biomarkers of tissue-specific diseases in readily accessible body fluids (e.g., biomarkers of a neurological disease (e.g., Alzheimer's disease antigen), to detect infection (especially detection of low titer latent viruses such as HIV), to detect fetal antigens in maternal blood, and to detect exogenous compounds (e.g., drugs or contaminants) in a subject's blood.

[0493] One potential source of the biomarker is also indicated in the table (e.g., "CSF"; cerebrospinal fluid). In many cases, the subject biosensors can detect those biomarkers in a different bodily fluid than that indicated. For example, biomarkers present in CSF can be identified in urine, blood, or saliva. It will also be clear to one of ordinary skill in the art that the CROF device of the present invention can be configured to capture and detect more biomarkers known in the art for diagnosing a disease or health condition.

[0494] Unless otherwise specified, a biomarker can be a protein or nucleic acid (e.g., mRNA) biomarker. Unless otherwise specified, a diagnosis can be associated with an increase or decrease in the level of a biomarker in a sample. A list of biomarkers, including diseases that can be used for diagnosis and samples in which the biomarkers can be detected, is described in Tables 1 and 2 of U.S. Provisional Patent Application No. 62 / 234,538, filed on September 29, 2015, and is incorporated herein by reference.

[0495] In some cases, the devices, systems and methods of the present invention are used to inform the health status of the person from whom the sample was taken. Health conditions that can be diagnosed or measured by the devices, systems and methods, devices and systems of the present invention include, but are not limited to: chemical balance; nutritional health; exercise; fatigue; sleep; stress; prediabetes; allergies; aging; exposure to environmental toxins, pesticides, herbicides, synthetic hormone analogs; pregnancy; menopause; and menopause. Table 3 of U.S. Provisional Patent Application 62 / 234,538 filed on September 29, 2015 provides a list of biomarkers that can be detected using the present CROF device (when used in conjunction with appropriate monoclonal antibodies, nucleic acids or other capture agents) and their associated health conditions.

[0496] J) Kit

[0497] As described above, aspects of the present invention include kits used in devices, systems and methods for performing the present invention. In certain embodiments, the kit includes instructions for practicing the method using a handheld device such as a mobile phone. These instructions may be present in the kit in a variety of forms, one or more of which may be present in the kit. One form that may appear in these instructions is information printed on a suitable medium or substrate, for example, in the packaging of the kit, in a package insert, on a sheet of paper with information printed on it. The device will be a computer-readable medium, such as a disk, CD, DVD, Blu-ray, computer-readable memory, etc., on which information has been recorded or stored. Another way is that it may be to use a website address that can access information from a deleted website via the Internet. The kit may further include software provided on a computer-readable medium for implementing a method for measuring an analyte on a device as described herein. There may be any convenient method in the kit.

[0498] In certain embodiments, the kit includes a detection agent including a detectable label, such as a fluorescently labeled antibody or oligonucleotide that specifically binds to the target analyte, for labeling the target analyte. The detection agent can be provided in a separate container as a CROF device, or it can be provided in a CROF device.

[0499] In certain embodiments, the kit includes a control sample, which includes a known detectable amount of an analyte to be detected in the sample. The control sample can be provided in a container and can be present in a solution at a known concentration, or can be provided in a dry form, such as lyophilized or freeze-dried. If provided in a dry form, the kit can also include a buffer for dissolving the control sample.

[0500] example

[0501] A) Example 1

[0502] The OAC is a QMAX device with two boards.

[0503] The first plate is a rectangular PMMA plate having a flat surface and a thickness in the range of 0.8 mm to 1.1 mm, 0.5 mm to 1.5 mm or 0.3 mm to 2 mm; a length in the range of 28 mm to 32 mm, 25 mm to 35 mm or 20 mm to 50 mm; and a width of 20 mm to 28 mm, 15 mm to 34 mm or 10 mm to 40 mm.

[0504] The second plate is a rectangular PMMA film having a flat surface and an array of micropillar arrays embossed on the flat surface. The thickness of the PMMA film is in the range of 0.8 mm to 1.1 mm, 0.5 mm to 1.5 mm, or 0.3 mm to 2 mm. The length is in the range of 28 mm to 32 mm, 25 mm to 35 mm, or 20 mm to 50 mm; the width is 20 mm to 28 mm, 15 mm to 34 mm, or 10 mm to 40 mm. In some embodiments, when the first plate and the second plate are placed together to hold the sample, at least three sides of the second plate are within the area of ​​the first plate. The pillar array has a rectangular or square shape with a flat top, the lateral dimension of the pillar is in the range of 30μm to 40μm, 25μm to 45μm, 20μm to 50μm, 10μm to 60μm, or 5μm to 70μm, the pillar height is in the range of 10μm to 30μm, 5μm to 40μm, 1μm to 50μm or 0.1μm to 100μm, and the distance between the centers of two adjacent pillars is 80 to 110μm, 60 to 130μm or 30 to 180μm or 30 to 200μm.

[0505] B) Hemoglobin measurement using OAC - using one wavelength

[0506] In the present experiment, the OAC is a QMAX device with two plates. The first plate is a 1mm thick flat PMMA substrate with dimensions of 30mm x 24mm. The second plate is a 175μm thick PMMA film with a micropillar array on it with dimensions of 24mm x 22mm. The pillar array has pillar dimensions of 30μm x 40μm, a pillar-to-pillar edge distance of 80μm, and pillar heights of 10μm or 30μm.

[0507] The sample is fresh whole blood (2.5uL for a 10μm column height and 5uL for a 30μm column height), which is dropped on the position of the first plate and then pressurized with the second plate.

[0508] In optical measurements, such as Figure 5 As shown, LED light is filtered by a bandpass filter (532nm to 576nm) and shines onto two 45-degree mirrors. The light then passes through a translucent diffuser to eliminate the coherence of the point source wavefront and ensure that intensity variations are only due to absorption. Finally, the light transmits the QMAX device and is collected by a lens and camera.

[0509] The LED lights and cameras used here can both come from mobile phones.

[0510] The pictures taken by the camera show that there are 2 areas. One area is the pillar area and the other area is the blood area.

[0511] The light absorption in the pillar region is negligible. Similarly, in the wavelength range of 532nm to 576nm, the extinction coefficients of oxygenated hemoglobin [HbO2] and deoxygenated hemoglobin [Hb] are similar ε Hb ≈ε HbO2 =44000-48000cm -1 / M.

[0512] therefore,

[0513] like Figure 6 As shown, I is the average intensity of the blood area, and Io is the average intensity of the center of the column area. When calculating the average intensity, we subtracted the 5 μm area near the column boundary to reduce the analysis error.

[0514]

[0515] We measured hemoglobin in blood ranging from 6 g / dL to 11 g / dL using the QMAX device setup and the commercially available Abbott Emerald blood cell counter and compared the results, as Figure 7 For each concentration, we measured 3 cards to calculate the standard deviation.

[0516] From the results, it can be seen that the repeatability (CV) of hemoglobin in the same blood sample measured by the QMAX card is about 5%, and the R2 value is 96% compared with the gold standard.

[0517] C) Example 3

[0518] Figure 8 An example of a hemoglobin measurement is shown, where an image of light transmission through a thin layer of whole blood (without lysis) in an OAC (e.g., the sample holder described in FIG1 ), and the light source is a diffuse light source (e.g., light diffusion is placed in front of a point light source), and the image is then captured by an iPhone. Figure 8 In the present invention, the light guide spacers are periodically placed on the QMAX card with a vertical periodic distance of 120 μm and a horizontal periodic distance of 110 μm.

[0519] Fig. 9 Shows the use of Figure 8 Example of image selection for determining the sample and reference areas for hemoglobin. Mark the boundaries of the sample area and reference.

[0520] exist Fig. 9 In the figure, the reference area (outer boundary is blue) is the inside of the light guide gasket. D, the distance between the edge of the reference area and the light guide gasket is 10μm; d, the distance between the edge of the sampling area and the light guide gasket is 30μm; T, the distance between the edge of the sampling area and the reference area is 40μm.

[0521] D) Image Processing

[0522] According to the present invention, the image processing algorithm for hemoglobin absorption measurement comprises the following steps:

[0523] 1. Light guide gasket detection

[0524] 2. Determination of reference area and sampling area

[0525] 3. Calculation of individual regions

[0526] 4. Merge

[0527] Light guide shim detection is used to detect and locate light guide shims that are periodically placed on the QMAX card. Various object detection algorithms can be used, including but not limited to template matching, blob detection, contour detection, etc. Detection can be performed in a single color channel in a color space (RGB, HSV, HSI, Lab, YCrCb, etc.), such as the green channel in the RGB color space, or the hue channel in the HSV space, or a combination of two or more color channels, such as using the red, green, and blue channels in the RGB color space, etc.

[0528] After the periodic light-guiding space is detected and located, a reference area (inside the light-guiding spacer) and a sampling area are selected. The size of the reference area and the sampling area, as well as the distance between the edges of the light-guiding space, the reference area and the sampling area are disclosed in the present invention.

[0529] For the reference area and the sampling area, they can be associated by relative position and distance. When one (or more) reference areas are associated with one (or more) sampling areas, hemoglobin absorption measurement values ​​can be calculated by the method disclosed in the present invention. One embodiment is to associate a reference area with the sampling area with the shortest distance, and calculate the hemoglobin absorption measurement value for each association.

[0530] The merging algorithm is to merge the hemoglobin absorption measurement values ​​of each sampling area and generate a final hemoglobin absorption measurement value. Various merging algorithms can be used, such as median, mean, maximum, minimum, k-means, etc.

[0531] In some embodiments, the imaging processing uses artificial intelligence and / or machine learning. In some embodiments, the imaging processing uses deep learning.

[0532] E) Using two wavelengths

[0533] Similar to the above experimental setup, except that 2 different bandpass filters were used and 2 pictures were taken.

[0534] Take a picture and calculate the blood at two wavelengths λ1 and λ2 For example, for 660nm and 940nm:

[0535]

[0536] We get:

[0537]

[0538] ε is the extinction coefficient of hemoglobin, [Hb] and [HbO2] are the concentrations of hemoglobin, and L is the optical path length through the sample or the gap size of a QMAX device.

[0539] Therefore, total hemoglobin concentration = [HbO2] + [Hb].

[0540] This method can further provide detailed information on the ratio of [HbO2] to [Hb].

[0541] F) Light guide gasket, sampling area and reference area

[0542] In some embodiments, the sampling area border has a size of 120 μm by 110 μm. The edge of the sampling area has a size of 60 μm by 45 μm; the size of the light guide spacer or column is 40 μm by 30 μm; the size of the reference area is 20 μm by 15 μm. In some embodiments, the area of ​​the reference area is 1 / 2 of the area of ​​the light guide spacer, the distance between the edge of the sampling area and the edge of the light guide spacer is 1 / 2 of the light guide spacer, and the area of ​​the sampling area is equal to the periodic distance of the spacer.

[0543] 1. A method or apparatus according to any one of the preceding claims, wherein the spacer has a cylindrical shape and a nearly uniform cross-section.

[0544] 2. A method or apparatus according to any one of the preceding claims, wherein the inter-spacer distance (SD) is equal to or less than about 150 μm (micrometers).

[0545] 3. A method or apparatus according to any one of the preceding claims, wherein the inter-spacer distance (SD) is equal to or less than about 100 μm (micrometers).

[0546] 4. A method or apparatus according to any one of the preceding claims, wherein the fourth power of the inter-shim distance (ISD) divided by the thickness (h) and Young's modulus (E) of the flexible board (ISD 4 / (hE)) is less than 5x10 6 μm 3 / GPa.

[0547] 5. A method or apparatus according to any one of the preceding claims, wherein the fourth power of the inter-shim distance (ISD) divided by the thickness (h) and Young's modulus (E) of the flexible board (ISD 4 / (hE)) is less than 5x10 5 μm 3 / GPa.

[0548] 6. The method or apparatus of any of the preceding claims, wherein the spacers have a columnar shape, a substantially flat top surface, a predetermined substantially uniform height, and a predetermined constant spacing interval that is at least about 2 times greater than the size of the analyte, wherein the Young's modulus of the spacer multiplied by the packing factor of the spacer is equal to or greater than 2 MPa, wherein the packing factor is the ratio of the spacer contact area to the total plate area, and wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1 (one).

[0549] 7. The method or apparatus of any of the preceding claims, wherein the spacers have a columnar shape, a substantially flat top surface, a predetermined substantially uniform height, and a predetermined constant spacing distance, the spacing distance being at least about 2 times greater than the size of the analyte, wherein the Young's modulus of the spacer multiplied by the packing factor of the spacer is equal to or greater than 2 MPa, wherein the packing factor is the ratio of the spacer contact area to the total plate area, and wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1 (one), wherein the fourth power of the spacer spacing (ISD) divided by the thickness (h) and the Young's modulus (E) of the flexible plate (ISD 4 / (hE)) is less than 5x10 6 μm 3 / GPa.

[0550] 8. The device of claim 1, wherein a ratio of a spacing distance of the spacers to an average width of the spacers is 2 or greater, and a filling factor of the spacers multiplied by a Young's modulus of the spacers is greater than 2 MPa.

[0551] 9. A method or apparatus according to any one of the preceding claims, wherein the analyte is an analyte in the detection of 5 of proteins, peptides, nucleic acids, synthetic compounds and inorganic compounds.

[0552] 10. A method or device according to any of the preceding claims, wherein the sample is selected from amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma or serum), breast milk, cerebrospinal fluid (CSF), cerumen, chyle, chyme, endolymph, peripheral lymph, feces, breath, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, exhaled breath condensate, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus and urine.

[0553] 11. A method or apparatus according to any preceding claim, wherein the spacer has a columnar shape and the ratio of the width to the height of the column is equal to or greater than one.

[0554] 12. A method according to any preceding claim, wherein the volume of sample deposited on one or both plates is unknown.

[0555] 13. A method or apparatus according to any preceding claim, wherein the spacer has a cylindrical shape and the cylindrical portion has a substantially uniform cross-section.

[0556] 14. A method or apparatus according to any one of the preceding claims, wherein the sample is used for the detection, purification and quantification of compounds or biomolecules associated with the stage of certain diseases.

[0557] 15. A method or apparatus according to any preceding claim, wherein the sample is related to infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, psychiatric diseases, neuropsychiatric diseases, lung diseases, kidney diseases and other and organic diseases.

[0558] 16. A method or apparatus according to any preceding claim, wherein the sample is relevant to the detection, purification or quantification of microorganisms.

[0559] 17. A method or apparatus according to any preceding claim, wherein the sample relates to viruses, fungi and bacteria from the environment, such as water, soil or a biological sample.

[0560] 18. A method or apparatus according to any preceding claim, wherein the sample is related to the detection, quantification of compounds or biological samples that pose a risk to food safety or national security (e.g., toxic waste, anthrax).

[0561] 19. A method or apparatus according to any preceding claim, wherein the sample relates to quantification of a vital parameter in a medical or physiological monitor.

[0562] 20. A method or apparatus according to any preceding claim, wherein the sample relates to glucose, blood, oxygen levels, total blood cell count.

[0563] 21. A method or apparatus according to any preceding claim, wherein the sample is concerned with the detection and quantification of specific DNA or RNA from a biological sample.

[0564] 22. A method or apparatus according to any preceding claim, wherein the sample is associated with sequencing and comparison of genetic sequences in the DNA in the chromosomes and mitochondria for genomic analysis.

[0565] 23. A method or apparatus according to any preceding claim, wherein the sample is relevant to the detection of a reaction product, for example during synthesis or purification of a drug.

[0566] 24. A method or apparatus according to any one of the preceding claims, wherein the sample is cells, tissue, body fluid and feces.

[0567] 25. The method or device according to any one of the preceding claims, wherein the sample is a sample for detecting proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds.

[0568] 26. A method or apparatus according to any one of the preceding claims, wherein the sample is a sample in the fields of human, veterinary, agricultural, food, environmental and pharmaceutical testing.

[0569] A method or apparatus according to any of the preceding claims, wherein the sample is selected from blood, serum, plasma, nasal swab, nasopharyngeal wash, saliva, urine, gastric fluid, spinal fluid, tears, feces, mucus, sweat, cerumen, oil, glandular secretions, cerebrospinal fluid, tissue, semen, vaginal fluid, tissue fluid derived from tumor tissue, eye fluid, spinal fluid, throat swab, breath, hair, nails, skin, biopsy, placental fluid, amniotic fluid, umbilical cord blood, lymph, cavity fluid, sputum, pus, microbiota, meconium, breast milk, exhaled condensate nasopharyngeal wash, nasal swab, throat swab, stool sample, hair, nails, ear wax, respiratory tract, connective tissue, muscle tissue, neural tissue, and other descriptions and additional examples of the present invention.

[0570] The present invention further includes the combination of the above disclosure and the various embodiments given below, which can be combined in various ways as long as the individual components do not contradict each other. The embodiments should be regarded as a single invention document: each document has other documents as references and is cited as a whole for all purposes, rather than as discrete independent documents. These embodiments include not only the disclosure in the current document, but also the documents cited, incorporated or claimed priority herein.

[0571] An apparatus for analyzing an analyte in a sample using light transmission, comprising:

[0572] The first board, the second board and the light guide spacer (LGS), wherein:

[0573] The first panel and the second panel are movable relative to each other into different configurations, including an open configuration and a closed configuration.

[0574] Each plate includes an inner surface having a sample contact area for contacting a sample containing or suspected of containing an analyte; and

[0575] Each light guide spacer has a cylindrical shape with a bottom end fixed to one of the plates and a top end having a flat surface, wherein the uniform height of the light guide spacers is less than 300 μm (micrometers), wherein, in a closed configuration, the top flat surface is in direct contact with the plates, and the cross-section of each light guide spacer is larger than the wavelength of light of the analyzed sample, wherein at least one guide spacer is located inside the sample;

[0576] Among them, the open configuration is that the two plates are separated, the interval between the two plates is not adjusted by the light-guiding spacer, and the sample is deposited on one or both plates.

[0577] wherein the closed configuration is the configuration configured in the open configuration after the sample is deposited; at least a portion of the sample is compressed by the two plates into a layer of uniform thickness, wherein the uniform thickness of the layer is limited by the t-plate and adjusted by the plates and the light guide spacer; and

[0578] An apparatus for analyzing an analyte in a sample using light transmission, comprising:

[0579] A first plate, a second plate, or more light guide spacers (LGS), a sampling area, and a reference area, wherein:

[0580] The first plate and the second plate are configured to sandwich a sample for optical transmission analysis by light in a thin layer between the plates, and each plate has a sample contact area on its inner surface that contacts the sample.

[0581] Each or more LGS has a cylindrical shape, is sandwiched between two plates, and both ends of the column directly contact one of the two plates forming the LGS plate contact area, and is configured to allow light to be transmitted from the first plate through the LGS to the second plate without passing through the sample,

[0582] The sampling region is a region through which light can sequentially pass through the first plate, the sample, and the second plate, wherein the sampling region does not have an LGS; and

[0583] The reference area is an area where light passes through the first plate, the light guide spacer, and the second plate in sequence without passing through the sample.

[0584] Among them, the LGS contact area and the LGS cross section are larger than the wavelength of light,

[0585] wherein the light guide spacer is surrounded by the sample or is surrounded by the sample, and

[0586] The thickness of the sample in the sampling area is less than 300 μm.

[0587] LGSs is a periodic array

[0588] LGSs are a periodic array and the spacing is less than 500 microns.

[0589] LGSs are a periodic array and the period is 250 μm or less.

[0590] LGSs are a periodic array and the period is less than 150 μm.

[0591] -Height less than 60μm

[0592] - Below 40 microns in height.

[0593] In a closed configuration: (a) the upper surface of at least one gasket in the sample contact area is in direct contact with one of the plates, and at least one gasket and the plate areas above and below at least one gasket define a reference area, wherein the reference area is transparent to light within a wavelength range, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate not occupied by a light-guiding gasket define a sampling area that is transparent to light within the same wavelength range.

[0594] An apparatus for analyzing an analyte in a sample using light transmission, comprising:

[0595] A first board, a second board and a light guide spacer, wherein:

[0596] The first plate and the second plate are configured to receive a sample containing or suspected of containing the analyte, wherein at least a portion of the sample is between and in contact with the two plates; and

[0597] The light guide spacer has a columnar shape and has a predetermined approximate height, wherein the light guide spacer is surrounded by the sample during light transmittance measurement.

[0598] Among them, the top surface and the bottom surface of the light guide gasket are basically flat, and the top surface and the bottom surface of the light guide gasket are in direct contact with the board.

[0599] wherein the area and average cross-section of the top and bottom surfaces of each spacer are respectively larger than the wavelength of light for analyzing the sample, and

[0600] wherein: (a) at least one spacer and areas of the plate directly above and below at least one spacer define a reference area that is transparent to light within a wavelength range that passes through the plate and spacer, and (b) at least one area in the sample contact area on one plate and at least one area in its corresponding area on the other plate are not occupied by a light-guiding spacer, thereby defining a sampling area that is transparent to light within the same wavelength range.

[0601] A device for analyzing an analyte in a sample, comprising:

[0602] A first board, a second board and a light guide spacer, wherein:

[0603] The first panel and the second panel are movable relative to each other into different configurations, including an open configuration and a closed configuration.

[0604] Each plate includes an inner surface having a sample contact area for contacting a sample containing or potentially containing an analyte; and

[0605] The light-guiding gasket has a cylindrical shape and a predetermined substantially uniform height, wherein the top and bottom surfaces of the light-guiding gasket are substantially flat and the bottom surface is fixed to the inner surface of one or more plates, wherein the top and bottom surfaces and the average cross-section of each gasket are respectively greater than 1 times the wavelength of light for analyzing a sample, and wherein at least one light-guiding gasket is within a sample contact area.

[0606] Among them, an open configuration is a configuration in which: the two plates are separated, the spacing between the plates is not limited by the light-guiding spacer, and the sample is deposited on one or both plates.

[0607] wherein the closed configuration is the configuration formed after the sample is deposited in the open configuration; in the closed state: at least a portion of the sample is compressed by the two plates into a highly uniform layer, wherein the uniform thickness of the layer is limited by the sample contact area of ​​the plates and adjusted by the plates and the light guide spacer; and

[0608] wherein, in a closed configuration, (a) at least one spacer in the sample contact region has its top surface in direct contact with one of the plates, and at least one spacer and at least one spacer region of the plates above and below define a reference region, wherein the reference region is transparent to light within a range of wavelengths, and (b) at least one region of the sample contact region on one plate and its corresponding region on the other plate is not occupied by a light-guiding spacer that defines a sampling region that is transparent to light within a range of wavelengths.

[0609] A device for analyzing t in a sample, comprising:

[0610] A first board, a second board and a light guide spacer, wherein:

[0611] The first plate and the second plate are configured to sandwich the sample in the thin layer.

[0612] A first board, a second board and a light guide spacer, wherein:

[0613] The first plate and the second plate are configured to sandwich the sample in the thin layer.

[0614] The cylindrical shape of the light-guiding spacer is sandwiched between two plates, wherein each end of the column directly contacts one of the plates, so that there is no sample between the end of the column and the corresponding plate. The guiding spacer is surrounded by or by the sample, and wherein the direct contact area and the average cross-section of the column are at least 1 μm, respectively. 2 (square micrometers) or larger.

[0615] The spacing between the inner surfaces of the plates is less than or equal to 200 μm.

[0616] An apparatus for sample analysis using light transmission, comprising:

[0617] The device of any one of the prior device embodiments, a light source, a camera; and an adapter, wherein

[0618] The light source is configured to emit light within a range of wavelengths configured to pass through the reference region.

[0619] The camera is configured to image the reference region and the sampling region.

[0620] The adapter is configured to position the device, the light source, and the camera relative to each other such that light from the light source passes through the reference region and the sampling region and is imaged by the camera.

[0621] The apparatus of any preceding apparatus embodiment, further comprising:

[0622] A processor is configured to process the image captured by the camera and determine a property of an analyte in the sample based on comparing light transmission from the reference region and the sampling region.

[0623] The device of any preceding claim, wherein the camera and the processor are part of a single mobile device.

[0624] The device of any preceding claim, wherein the light source and the processor are part of a single mobile device.

[0625] The device of any preceding claim, wherein said light source, said camera and said processor are part of a single mobile device.

[0626] The device of any previous device embodiment, wherein the mobile device is a smartphone.

[0627] Using transmitted light for a method for sample analysis, comprising the steps of:

[0628] An apparatus as in any preceding apparatus embodiment;

[0629] depositing a sample in an open configuration of the device, wherein the sample may contain an analyte;

[0630] Put the device into a closed state;

[0631] having a light source having a wavelength configured to pass through a reference region of the device;

[0632] having an imager configured to image a reference region and a sampling region of the device;

[0633] An adapter is provided that is configured to position the device, the light source, and the camera relative to each other so that light from the light source passes through the reference region and the sampling region and is imaged by the camera.

[0634] The nature of the analyte is determined by comparing the light transmittance from the sample area and the reference area.

[0635] The apparatus, method or system of any preceding claim, wherein the analyte is haemoglobin.

[0636] The apparatus, method or system of any preceding claim, wherein the analyte is a cell.

[0637] All analytes were added.

[0638] Add different applications.

[0639] Increase the critical distance between light transmissions.

[0640] The apparatus, method or system of any preceding claim, wherein the thickness of said sample layer is adjusted by the plate and said light-guiding spacer and is substantially the same as the uniform height of said light-guiding spacer;

[0641] The apparatus, method or system of any preceding claim, wherein the analyte is red blood cells.

[0642] The apparatus, method or system of any preceding claim, wherein the analyte is a leukocyte.

[0643] The apparatus, method or system of any preceding claim, wherein the reference area and the sampling area are of the same size.

[0644] The apparatus, method or system of any preceding claim, wherein the reference region is within a corresponding region of a cross-section of the light guide spacer.

[0645] The apparatus, method or system of any preceding claim, wherein the reference area is less than 0.1 μm^2, less than 0.2 μm^2, less than 0.5 μm^2, less than 1 μm^2, less than 2 μm^2, less than 5 μm^2, less than 10 μm^2, less than 20 μm^2, less than 50 μm^2, less than 100 μm^2, less than 200 μm^2, less than 500 μm^2, less than 1000 μm^2, less than 2000 μm^2, less than 5000μm^2, less than 10000μm^2, less than 20000μm^2, less than 50000μm^2, less than 100000μm^2, less than 200000μm^2, less than 500000μm^2, less than 1mm^2, less than 2mm^2, less than 5mm^2, less than 10mm^2, less than 20mm^2, or less than 50mm^2, or in a range between two values.

[0646] The apparatus, method or system of any preceding claim, wherein the device further comprises a plurality of light-guiding spacers having a substantially uniform height, and wherein at least one of the light-guiding spacers is within the sample contact area.

[0647] The apparatus, method or system of any preceding claim, wherein the device further comprises a plurality of light-guiding spacers having a substantially uniform height, wherein a distance between two adjacent light-guiding spacers is known, and wherein at least one light-guiding spacer is located within the sample contact area.

[0648] The apparatus, method or system of any preceding claim, wherein the device further comprises a plurality of light-guiding spacers having a substantially uniform height, wherein the distance between two adjacent light-guiding spacers is known and substantially constant (i.e., the light-guiding spacers are substantially a periodic array), and wherein at least one of the light-guiding spacers is within the sample contact area.

[0649] The apparatus, method or system of any preceding claim, wherein the bottom surface of the light guide spacer is secured to the inner surface of one of the plates by moulding the light guide spacer on the inner surface of the plate.

[0650] The apparatus, method or system of any preceding claim, wherein the bottom surface of the light-guiding spacer is secured to an inner surface of one of the plates and is made of the same material as the inner surface.

[0651] The apparatus, method or system of any preceding claim, wherein the bottom surface of the light-guiding spacer is secured to the inner surface of one of the plates and is made of the same material as the inner surface and the bottom surface of the bottom plate. The light-guiding spacer has no interface with the inner surface of the plate.

[0652] The apparatus, method or system of any preceding claim, wherein the wavelength of the light is greater than 300 nm, and wherein the wavelength of the light is also less than 20 μm, less than 15 μm, less than 10 μm, less than 20 nm, less than 5 μm, less than 4 μm, less than 3 μm, less than 2 μm, less than 1 μm, less than 800 nm, less than 750 nm, less than 700 nm, less than 650 nm, less than 600 nm, less than 550 nm, less than 500 nm, less than 450 nm, less than 400 nm, or a range between two values.

[0653] The apparatus, method or system of any preceding claim, wherein the wavelength of the light is greater than 500 nm, and wherein the wavelength of the light is also less than 600 nm, less than 590 nm, less than 580 nm, less than 100 nm, less than 570 nm, less than 560 nm, less than 550 nm, less than 540 nm, less than 530 nm, less than 520 nm, less than 510 nm, or a range between two values.

[0654] The apparatus, method or system of any preceding claim, wherein the average cross-section of each light-guiding spacer is less than 1 μm^2 (square micrometer), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2, 10,000 μm^2, 30,000 μm^2, 100,000 μm^2, 200,000 μm^2, 500,000 μm^2, 1 mm^2, 2 mm^2, 5 mm^2, 10 mm^2, 50 mm^2 or a range between two of the following values.

[0655] The apparatus, method or system of any preceding claim, wherein the average cross-section of each light-guiding spacer is less than 1 μm^2 (square micrometer), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2, 10,000 μm^2, 30,000 μm^2, 100,000 μm^2, 200,000 μm^2 or a range between two values.

[0656] The apparatus, method or system of any preceding claim, wherein the average cross-section of each light-guiding spacer is less than 1 μm^2 (square micrometer), 10 μm^2, 20 μm^2, 30 μm^2, 50 μm^2, 100 μm^2, 150 μm^2, 200 μm^2, 300 μm^2, 500 μm^2, 1000 μm^2, 2000 μm^2, 5000 μm^2, 10,000 μm^2, 30,000 μm^2 or a range between two values.

[0657] The apparatus, method or system of any preceding claim, wherein the sample contact area is greater than 100 μm^2 (square micrometers), greater than 200 μm^2, greater than 400 μm^2, greater than 600 μm^2, greater than 800 μm^2, greater than 1,000 μm^2, greater than 2,000 μm^2, greater than 4,000 μm^2, greater than 6,000 μm^2, greater than 8,000 μm^2, greater than 10,000 μm^2, greater than 20,000 μm^2, greater than 40,000 μm^2, greater than 60,000 μm^2, greater than 80,000 μm^2, greater than 100,000 μm^2, greater than 200,000 μm^2, greater than 250,000 μm^2, greater than 500,000 μm^2 (square micrometers), or within a range between two values.

[0658] The apparatus, method or system of any preceding claim, wherein the predetermined constant spacing interval is at least about 2 times greater than the size of the analyte.

[0659] The apparatus, method or system of any preceding claim, wherein the predetermined constant spacing interval is at least 2 times, at least 6 times, at least 8 times, at least 10 times, at least 20 times, at least 40 times, at least 60 times, at least 80 times or at least 100 times larger than the size of the analyte.

[0660] The apparatus, method or system of any preceding claim, wherein the analyte is a biological marker, an environmental marker or a food marker.

[0661] The device, method or system of any preceding claim, wherein the analyte is a biomarker indicative of the presence or severity of a disease or condition.

[0662] The apparatus, method or system of any preceding claim, wherein the analyte is a cell, a protein or a nucleic acid.

[0663] The apparatus, method or system of any preceding claim, wherein the analyte is haemoglobin.

[0664] The apparatus, device or method of any preceding claim, wherein the analyte comprises proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles and other molecules, compounds, mixtures and substances thereof.

[0665] The apparatus, device or method of any preceding claim, wherein the sample is in raw, diluted or processed form of a body fluid, feces, amniotic fluid, aqueous body fluid, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, cerumen, chyle, bells, endolymph, perilimph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine or exhaled breath condensate.

[0666] The apparatus, device or method of any preceding claim, wherein the sample is in the form of raw, diluted or processed blood.

[0667] The apparatus, device or method of any preceding claim, wherein the sample comprises whole blood.

[0668] The apparatus, device or method of any preceding claim, wherein the sample comprises an aggregating agent which causes aggregation of the interfering elements.

[0669] The apparatus, device or method of any preceding claim, wherein the sample holder comprises a hole configured to hold a sample.

[0670] The apparatus, device or method of any preceding claim, wherein the sample holder comprises first and second plates and a spacer.

[0671] The apparatus, device or method of any preceding claim, wherein the sample holder comprises a first plate, a second plate and a spacer, wherein the spacer is configured to adjust the gap between the plates when the plates are pressed toward each other, thereby compressing the plates to press the sample into a thin layer.

[0672] The apparatus, device or method of any preceding claim, wherein the sample holder comprises a first plate, a second plate and a spacer, and wherein:

[0673] The panels are movable relative to each other into different configurations, including an open configuration and a closed configuration.

[0674] In the open state: the two plates are separated, the spacing between the two plates is not limited by the spacer, and the sample is deposited on one or both plates.

[0675] In the closed configuration, which is configured in the open configuration after sample deposition: at least a portion of the sample is compressed by the two plates into a highly uniform layer and substantially stagnant relative to the plates, wherein the layer is conditioned by the plates and the spacers.

[0676] The apparatus, device or method of any preceding claim, wherein the sample holder comprises a Q-card comprising a first plate, a second plate and a spacer, wherein the spacer is configured to adjust the gap between the plates when the plates are pressed against each other to compress the sample into a thin layer.

[0677] The apparatus, device or method of any preceding claim, wherein

[0678] The sample holder includes a first plate, a second plate, and a spacer, wherein the spacer has a uniform height and a constant spacing distance. and

[0679] ii. The sample is compressed by the sample holder into a thin layer of uniform thickness, which is adjusted by the height of the spacer.

[0680] Apparatus, means or method as claimed in any preceding claim, wherein the sample is compressed into a layer having a uniform thickness substantially equal to the uniform height of the spacers fixed to the one or both plates.

[0681] The apparatus, kit or method of any preceding claim, wherein the sample is pressed into a layer of uniform thickness, the thickness varying by less than 15%, 10%, 5%, 2%, 1% or within a range between any two values.

[0682] The apparatus, device or method of any preceding claim, wherein the sample, when compressed, has a thickness of less than 500 nm, less than 1000 nm, less than 2 μm, less than 5 μm, less than 10 μm, less than 20 μm, less than 50 μm, less than 100 μm, less than 150 μm, less than 200 μm, less than 300 μm, less than 500 μm, less than 800 μm, less than 1 mm (millimeter), less than 2 mm, less than 3 mm, less than 5 mm, less than 10 mm or in a range between two values.

[0683] The apparatus, device or method of any preceding claim, wherein the sample holder comprises a first plate and a second plate, wherein the thickness of each plate is less than or equal to 500 nm, less than or equal to 1000 nm, less than or equal to 2 μm, less than or equal to 5 μm, less than or equal to 10 μm, less than or equal to 20 μm, less than or equal to 50 μm, less than or equal to 100 μm, less than or equal to 150 μm, less than or equal to 200 μm, less than or equal to 300 μm, less than or equal to 500 μm, less than or equal to 800 μm, less than or equal to 1 mm (millimeter), less than or equal to 2 mm, less than or equal to 3 mm, less than or equal to 5 mm, less than or equal to 10 mm, or between any two of these values.

[0684] The apparatus, device or method of any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less, 0.1 μm or less, 50 nm or less, 20 nm or less, 10 nm or less, or a range between two values.

[0685] The apparatus, device or method of any preceding claim, wherein at least a portion of the sample is compressed into a thin layer, and wherein only interference-rich regions exist for a specific portion of the sample having an average thickness of 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less, 0.1 μm or less, 50 nm or less, 20 nm or less, 10 nm or less, or a range between two values.

[0686] The apparatus, device or method of any preceding claim, wherein at least a portion of the sample is compressed into a thin layer, wherein only interference-poor regions exist for a particular portion of the sample having an average thickness of 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less, 0.1 μm or less, 50 nm or less, 20 nm or less, 10 nm or less, or within a range of either value.

[0687] The apparatus, device or method of any preceding claim, wherein at least a portion of the sample is compressed to an average thickness of 0.5-2 μm, 0.5-3 μm, 0.5-5 μm, 0.5-10 μm, 0.5-20 μm, 0.5-30 μm or 0.5-50 μm.

[0688] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 500 μm.

[0689] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 200 μm.

[0690] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 100 μm.

[0691] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 50 μm.

[0692] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 25 μm.

[0693] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 10 μm.

[0694] 7. The device, apparatus or method of claim 1, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of 5 μm or less.

[0695] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 3 μm.

[0696] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 2 μm.

[0697] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of less than 1 μm.

[0698] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of 500 nm or less.

[0699] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness of 100 nm or less.

[0700] Apparatus, means or method as claimed in any preceding claim, wherein at least a portion of the sample is compressed into a thin layer having an average thickness in the range 0.5-2 μm, 0.5-3 μm or 0.5-5 μm.

[0701] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is in the range of 2 μm to 2.2 μm and the sample is blood.

[0702] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is in the range of 2.2 μm to 2.6 μm and the sample is blood.

[0703] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is in the range of 1.8 μm to 2 μm and the sample is blood.

[0704] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is in the range of 2.6 μm to 3.8 μm and the sample is blood.

[0705] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is in the range of 1.8 μm to 3.8 μm and the sample is whole blood, not diluted by another liquid.

[0706] The apparatus, device or method of any preceding claim, wherein the average thickness of the layer of uniform thickness is approximately equal to the smallest dimension of an analyte in the sample.

[0707] The apparatus, device or method of any preceding claim, wherein the final sample thickness device is configured to analyze the sample in 300 seconds or less.

[0708] The apparatus, device or method of any preceding claim, wherein the final sample thickness device is configured to analyze the sample in 180 seconds or less.

[0709] The apparatus, device or method of any preceding claim, wherein the final sample thickness device is configured to analyze the sample in 60 seconds or less.

[0710] The apparatus, device or method of any preceding claim, wherein the final sample thickness device is configured to analyze the sample in 30 seconds or less.

[0711] The apparatus, device or method of any preceding claim, wherein the imager comprises a camera.

[0712] The apparatus, device or method of any preceding claim, wherein the imager is part of the detector.

[0713] The apparatus, device or method of any preceding claim, wherein the imager is integral to the detector.

[0714] The apparatus, device or method of any preceding claim, wherein the imager is directed by the software to capture one or more images of the sample, identify the interfering element regions and the interfering element-free regions, and digitally distinguish the interfering element regions from the interfering element-free regions.

[0715] The apparatus, device or method of any preceding claim, wherein the imager comprises a filter configured to filter a signal from the sample.

[0716] The apparatus, device or method of any preceding claim, wherein the imager comprises a light source configured to illuminate the sample.

[0717] The apparatus, device or method of any preceding claim, wherein the apparatus or method is used to detect proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles and other molecules, compounds, mixtures and substances thereof.

[0718] The device, apparatus or method of any preceding claim, wherein the device or method is for use in the diagnosis, management and / or prevention of human diseases and conditions.

[0719] The apparatus, device or method of any preceding claim, wherein the apparatus or method is for use in diagnosing, managing and / or preventing veterinary diseases and conditions, or for use in diagnosing, managing and / or preventing plant diseases and conditions.

[0720] The apparatus, device or method of any preceding claim, wherein the apparatus or method is used for environmental testing and decontamination.

[0721] The device, apparatus or method of any preceding claim, wherein the device or method is for use in agricultural or veterinary applications.

[0722] The apparatus, device or method of any preceding claim, wherein the apparatus or method is for use in food testing.

[0723] The device, apparatus or method of any preceding claim, wherein the device or method is used for drug testing and prevention.

[0724] The device, apparatus or method of any preceding claim, wherein the device or method is for detecting and / or measuring an analyte in blood.

[0725] The device, apparatus or method of any preceding claim, wherein the device or method is for use in a colorimetric assay.

[0726] The device, apparatus or method of any preceding claim, wherein the device or method is for use in a fluorescence assay.

[0727] Apparatus, means or method as claimed in any preceding claim, wherein the plates are movable relative to each other.

[0728] The apparatus, device or method of any preceding claim wherein the spacers are secured to one or both of the plates and are of uniform height.

[0729] The apparatus, device or method of any preceding claim, wherein the first and second plates are configured to compress the sample into a layer of uniform thickness, the thickness being substantially equal to the height of the spacer.

[0730] The apparatus, device or method of any preceding claim, wherein the spacer has a uniform height of 1 mm or less, 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less, 0.1 μm or less, 50 nm or less, 20 nm or less, 10 nm or less, or a range between two values.

[0731] The apparatus, device or method of any preceding claim, wherein the spacers have a uniform height in the range of 0.5-2 μm, 0.5-3 μm, 0.5-5 μm, 0.5-10 μm, 0.5-20 μm, 0.5-30 μm or 0.5-50 μm.

[0732] The apparatus, device or method of any preceding claim, wherein at least one of the plates has a diameter of 100 mm or less, 50 mm or less, 25 mm or less, 10 mm or less, 5 mm or less, 1 mm or less, 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less or 0.1 μm or less, or in a range between two values.

[0733] The apparatus, device or method of any preceding claim, wherein at least one of the plates has a thickness in the range of 0.5 to 1.5 mm; around 1 mm; in the range of 0.15 to 0.2 mm; or about 0.175 mm.

[0734] The apparatus, device or method of any preceding claim, wherein at least one of the plates has a thickness of 1 mm 2 Below, 10mm 2 Below, 25mm 2 Below 50mm 2 Below, 75mm 2 Below, 1cm 2 (square centimeters) or less, 2cm 2 Below, 3cm 2 Below, 4cm 2 Below, 5cm 2 Below, 10cm 2 Below 100cm 2 Below 500cm 2 Below 1000cm 2 Below 5000cm 2 Below 10,000cm 2 Below 10,000cm 2 The lateral area below or in the range between two values.

[0735] The apparatus, device or method of any preceding claim, wherein at least one of the plates has a width of 500 mm. 2 Up to 1000mm 2 Within or about 750mm 2 The lateral area

[0736] The apparatus, device or method of any preceding claim, wherein the Young's modulus of the gasket multiplied by the packing factor of the gasket is equal to or greater than 10 MPa, wherein the packing factor is the ratio of the area of ​​the gasket contacted by the uniform thickness layer to the total plate area contacted by the uniform thickness layer.

[0737] The apparatus, device or method of any preceding claim, wherein the thickness of the flexible sheet multiplied by the Young's modulus of the flexible sheet is in the range of 60 to 750 GPa-μm.

[0738] The apparatus, device or method of any preceding claim, wherein, for a flexible board, the fourth power of the inter-pad distance (ISD) divided by the thickness (h) of the flexible board and the Young's modulus (E) of the flexible board, ISD 4 / (hE) is equal to or less than 10 6 μm 3 / GPa.

[0739] The apparatus, device or method of any preceding claim, wherein one or both plates include position markings on or in the surface of the plate, the position markings providing position information of the plate.

[0740] Apparatus, means or method as claimed in any preceding claim, wherein one or both plates include a scale on or within its surface which provides information on the lateral dimensions of the structure of the sample and / or the plate.

[0741] The apparatus, device or method of any preceding claim, wherein one or both plates include image markings on the surface or in the interior of the plate, said image markings facilitating imaging of the sample.

[0742] The apparatus, device or method of any preceding claim, wherein the distance between the spacers is in the range 7 μm to 120 μm.

[0743] The apparatus, device or method of any preceding claim, wherein the distance between the spacers is in the range 120 μm to 200 μm.

[0744] The apparatus, device or method of any preceding claim, wherein the spacer is a post having a cross-sectional shape selected from circular, polygonal, circular, square, rectangular, elliptical, oval or any combination thereof.

[0745] The apparatus, device or method of any preceding claim, wherein the spacers have a cylindrical shape and have a substantially flat top surface, wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.

[0746] The apparatus, device or method of any preceding claim, wherein the ratio of the lateral dimension of each spacer to its height is at least 1.

[0747] The apparatus, device or method of any preceding claim, wherein the smallest transverse dimension of the spacer is less than or substantially equal to the smallest dimension of the analyte in the sample.

[0748] The apparatus, device or method of any preceding claim, wherein the smallest lateral dimension of the spacer is in the range 0.5 μm to 100 μm.

[0749] The apparatus, device or method of any preceding claim, wherein the smallest lateral dimension of the spacer is in the range 0.5 μm to 10 μm.

[0750] The apparatus, device or method of any preceding claim, wherein the spacer has a columnar shape and the side wall corners of the spacer have a rounded shape with a radius of curvature of at least 1 μm.

[0751] The apparatus, device or method of any preceding claim, wherein the spacer has a thickness of at least 100 / mm 2 density.

[0752] The apparatus, device or method of any preceding claim, wherein the spacer has a thickness of at least 1000 / mm 2 density.

[0753] The apparatus, device or method of any preceding claim, wherein at least one of the plates is transparent.

[0754] The apparatus, device or method of any preceding claim, wherein at least one of the plates is made from a flexible polymer.

[0755] The apparatus, device or method of any preceding claim wherein the gasket is incompressible with respect to pressure compressing the plates and / or independently only one of the plates is flexible.

[0756] The apparatus, device or method of any preceding claim, wherein the flexible sheet has a thickness in the range 10 μm to 200 μm.

[0757] The apparatus, device or method of any preceding claim, wherein the variation in sample thickness is less than 30%.

[0758] The apparatus, device or method of any preceding claim, wherein the variation in sample thickness is less than 10%.

[0759] The apparatus, device or method of any preceding claim, wherein the variation in sample thickness is less than 5%.

[0760] The apparatus, device or method of any preceding claim, wherein the first and second panels are connected and configured to change from an open configuration to a closed configuration by folding the panels.

[0761] The apparatus, device or method of any preceding claim, wherein the first and second panels are connected by a hinge and are configured to change from an open configuration to a closed configuration by folding the panels along the hinge.

[0762] The apparatus, device or method of any preceding claim, wherein the first and second panels are connected to the panels by hinges as separate materials and are configured to change from an open configuration to a closed configuration by folding the panels along the hinges.

[0763] The apparatus, device or method of any preceding claim, wherein the first and second panels are formed from a single piece of material and are configured to change from an open configuration to a closed configuration by folding the panels.

[0764] The apparatus, device or method of any preceding claim, wherein the sample layer of uniform thickness is at least 1 mm 2 It is uniform over the lateral area.

[0765] Apparatus, device or method as claimed in any preceding claim, wherein the spacers are secured to the plate by direct rolling of the plate or injection moulding of the plate.

[0766] The apparatus, device or method of any preceding claim, wherein the material of the plates and the spacers is selected from polystyrene, PMMA, PC, COC, COP or another plastic.

[0767] The apparatus, method or system of any preceding claim, wherein the first plate further comprises a scattering surface on an inner surface thereof.

[0768] The apparatus, method or system of any preceding claim, wherein said first sheet is substantially transparent.

[0769] The apparatus, method or system of any preceding claim, wherein the reflectivity of the first plate is in the range 1% to 80%.

[0770] The apparatus, method or system of any preceding claim, wherein the reflectivity of the first plate is greater than 1%, 2%, 4%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70% or 80%, or a range between two values.

[0771] The apparatus, method or system of any preceding claim, wherein the first plate is made of PMMA.

[0772] The apparatus, method or system of any preceding claim, wherein the reflectivity of the second plate is in the range 1% to 100%.

[0773] The apparatus, method or system of any preceding claim, wherein the reflectivity of the second plate is greater than 1%, 2%, 4%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95%, or a range between two values.

[0774] The apparatus, method or system of any preceding claim, wherein at least a portion of the inner and outer surfaces of the first plate are substantially planar.

[0775] The apparatus, method or system of any preceding claim, wherein the scattering surface has a reflectivity greater than 90% across the entire spectrum of illumination light used to illuminate the sample.

[0776] The apparatus, method or system of any preceding claim, wherein the scattering surface has a reflectivity greater than 50%, 60%, 70%, 80%, 90% or in a range between two values.

[0777] The apparatus, method or system of any preceding claim, wherein the scattering surface is coated with a metallic film.

[0778] The apparatus, method or system of any preceding claim, wherein the scattering surface is coated with a metal film and the thickness of the metal film is in the range of 10 nm to 100 nm.

[0779] The apparatus, method or system of any preceding claim, wherein the scattering surface is coated with a metal film and the thickness of the metal film is less than 10 nm, 15 nm, 20 nm, 25 nm, 30 nm, 40 nm, 50 nm, 60 nm, 80 nm or 100 nm, or a range between two values.

[0780] The apparatus, method or system of any preceding claim, wherein the scattering surface is coated with a metal film, and the metal film comprises one or more of an aluminum film, a silver film and a gold film.

[0781] The apparatus, method or system of any preceding claim, said scattering surface having a transmittance in the range 10% to 30%.

[0782] The apparatus, method or system of any preceding claim, wherein the scattering surface has a transmissivity of less than 10%, 15%, 20%, 25% or 30%, or within a range between two values.

[0783] The apparatus, method or system of any preceding claim, wherein the scattering surface of one of the first and second plates and the other plate enhances light capture.

[0784] The apparatus, method or system of any preceding claim, wherein the scattering surface has a reflectivity that is Lambertian-dominated in at least one wavelength range.

[0785] The apparatus, method or system of any preceding claim, wherein the scattering surface has a Lambertian reflectivity greater than 0.8 in the at least one wavelength range.

[0786] The apparatus, method or system of any preceding claim, wherein the scattering surface is a surface formed by chemical etching.

[0787] The apparatus, method or system of any preceding claim, wherein the scattering surface is a surface formed by nanoimprint lithography.

[0788] The apparatus, method or system of any preceding claim, wherein said scattering surface covers substantially all of the sample contact area of ​​said first plate.

[0789] The apparatus, method or system of any preceding claim, wherein the scattering surface covers a portion of a sample contacting area of ​​the first plate.

[0790] The apparatus, method or system of any preceding claim, wherein said portion of said first plate is substantially transparent and covers substantially all of the sample contact area of ​​said second plate.

[0791] An apparatus, method or system as claimed in any preceding claim, wherein said portion of said first plate is substantially transparent and covers a portion of the sample contacting area of ​​said second plate.

[0792] The apparatus, method or system of any preceding claim, wherein the scattering surface comprises a non-smooth surface that is concave, convex and wavy.

[0793] The apparatus, method or system of any preceding claim, wherein the scattering surface comprises a periodic texture.

[0794] The apparatus, method or system of any preceding claim, wherein the scattering surface comprises a non-periodic texture.

[0795] The apparatus, method or system of any preceding claim, wherein the scattering surface has an average roughness in the range between 2 μm and 5 μm.

[0796] The apparatus, method or system of any preceding claim, wherein the average roughness of the scattering surface is less than 2.0 μm, 2.5 μm, 3.0 μm, 3.5 μm, 4.0 μm, 4.5 μm or 5.0 μm, or within a range between two values.

[0797] The apparatus, method or system of any preceding claim, wherein the light-guiding spacer is secured to the inner surface of the first plate and has a predetermined uniform height.

[0798] The apparatus, method or system of any preceding claim, wherein the adapter comprises:

[0799] an adapter housing having an exit aperture for positioning the imager;

[0800] Passive illuminators; and

[0801] Therein, the passive illuminator is located on the adapter and positioned around the periphery of the exit aperture.

[0802] The apparatus, method or system of any preceding claim, wherein the adapter housing is configured to reduce the entry of ambient light external to the adapter housing into the interior of the adapter housing.

[0803] The apparatus, method or system of any preceding claim, wherein the adapter comprises one or two light guides, one end of each light guide being aligned with the entrance of the optical chamber so that light entering that end of the light guide passes through the light guide to the corresponding end of the passive illuminator.

[0804] Apparatus, method or system as claimed in any preceding claim, wherein the one or more light guides and the passive illuminator are jointly formed from optical fibres.

[0805] The apparatus, method or system of any preceding claim, wherein the passive illuminator is in the form of a ring configured to surround an optical axis of a lens in a camera of the smartphone when the device is engaged with the smartphone.

[0806] The apparatus, method or system of any preceding claim, wherein the device further comprises an auxiliary lens, the optical axis of the auxiliary lens being aligned with the optical axis of a lens in a camera of the smartphone when the device is engaged with the smartphone.

[0807] The device, method or system of any preceding claim, wherein the device further comprises an auxiliary lens having a diameter of at least 2 mm, 3 mm, 4 mm, 5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 40 mm or 50 mm, or a range between two values.

[0808] The apparatus, method or system of any preceding claim, wherein the device further comprises a light concentrator configured to be placed in front of a light source of the smartphone when the device is engaged with the smartphone.

[0809] The apparatus, method or system of any preceding claim, wherein the diffuser comprises a volume diffusing material.

[0810] The apparatus, method or system of any preceding claim, wherein the diffuser comprises at least one textured surface.

[0811] The apparatus, method or system of any preceding claim, wherein the diffuser comprises a diffuser plate of substantially uniform thickness.

[0812] The apparatus, method or system of any preceding claim, wherein the diffuser comprises a diffuser plate having a thickness greater than an average thickness of the diffuser plate.

[0813] The apparatus, method or system of any preceding claim, further comprising a reflector configured to reflect light emitted from the passive illuminator towards the diffuser.

[0814] Apparatus, method or system as claimed in any preceding claim, wherein the passive illuminator is formed by side illuminating fibres.

[0815] The apparatus, method or system of any preceding claim, wherein the passive illuminator is rotationally symmetric.

[0816] The apparatus, method or system of any preceding claim, wherein the passive illuminator is circular.

[0817] The apparatus, device or method of any preceding claim, wherein the passive illuminator is circular with a diameter in the range 5 mm to 100 mm.

[0818] The apparatus, device or method of any preceding claim, wherein the passive illuminator is circular and has a diameter of at least 5 mm, 10 mm, 15 mm, 20 mm, 25 mm, 30 mm, 40 mm, 50 mm, 60 mm, 80 mm, or 100 mm, or a range therebetween.

[0819] The apparatus, device or method of any preceding claim, wherein the passive illuminator is in the form of a convex polygon.

[0820] The apparatus, device or method of any preceding claim, wherein the passive illuminator is in the form of a star-shaped polygon.

[0821] The apparatus, device or method of any preceding claim, wherein the optical axis of the lens passes through the centre of the passive illuminator.

[0822] The apparatus, device or method of any preceding claim, wherein the passive illuminator is rotationally asymmetric.

[0823] The apparatus, device or method of any preceding claim, wherein the passive illuminator is elliptical.

[0824] The apparatus, device or method of any preceding claim, wherein the passive illuminator has a substantially uniform cross-section.

[0825] The apparatus, device or method of any preceding claim, wherein all cross-sections of the passive illuminator over more than 50% of its length are of substantially the same shape.

[0826] The apparatus, device or method of any preceding claim wherein substantially all of the cross-sections are circular in shape.

[0827] The apparatus, device or method of any preceding claim wherein substantially all of the cross-sections are circular in shape and have a diameter in the range 1.0 mm to 3.0 mm.

[0828] The apparatus, device or method of any preceding claim, wherein substantially all of the cross-sectional shapes have a diameter of at least 0.5 mm, 1.0 mm, 1.5 mm, 2.0 mm, 2.5 mm, 3 mm, 4 mm, 5 mm, 6 mm, 8 mm, or 10 mm, or a range therebetween.

[0829] The apparatus, device or method of any preceding claim wherein substantially all of the cross-sections are elliptical in shape.

[0830] The apparatus, device or method of any preceding claim, wherein at least a portion of the side wall of the passive illuminator is formed by a diffusing surface.

[0831] The apparatus, device or method of any preceding claim, wherein the sample is in raw, diluted or processed form of a body fluid, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, cerumen, chyle, bells, endolymph, perilimph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, rhubarb, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine or exhaled breath condensate.

[0832] The apparatus, method or system of any preceding claim, wherein the sample is in the form of raw, diluted or processed blood.

[0833] The apparatus, method or system of any preceding claim, wherein the sample comprises whole blood.

[0834] The apparatus, method or system of any preceding claim, wherein the sample is a biological sample, a chemical sample, an environmental sample or a food sample.

[0835] The apparatus, method or system of any preceding claim, wherein the analyte is a biological marker, an environmental marker, or a food marker.

[0836] The device, method or system of any preceding claim, wherein the analyte is a biomarker indicative of the presence or severity of a disease.

[0837] The apparatus, method or system of any preceding claim, wherein the analyte is a cell, a protein or a nucleic acid.

[0838] The apparatus, method or system of any preceding claim, wherein the analyte comprises proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles, and other molecules, compounds, mixtures and substances thereof.

[0839] The apparatus, method or system of any preceding claim, wherein the sample holder comprises a hole for holding the sample.

[0840] The apparatus, method or system of any preceding claim, wherein the sample holder comprises first and second plates and a spacer.

[0841] The apparatus, method or system of any preceding claim, wherein the sample holder comprises a first plate, a second plate and a spacer, wherein the spacer is configured to adjust the gap between the plates when the plates are pressed against each other, so that the sample is compressed into a thin layer when the plates are compressed.

[0842] The apparatus, method or system of any preceding claim, wherein the sample holder comprises a first plate, a second plate and a spacer, and wherein:

[0843] The panels are movable relative to each other into different configurations, including an open configuration and a closed configuration.

[0844] In the open state: the two plates are separated, the spacing between the two plates is not limited by the spacer, and the sample is deposited on one or both plates.

[0845] In the closed configuration, which is configured in the open configuration after sample deposition: at least a portion of the sample is compressed by the two plates into a highly uniform layer and substantially stagnant relative to the plates, wherein the layer is conditioned by the plates and the spacers.

[0846] The apparatus, method or system of any preceding claim, wherein the sample holder comprises a Q-card comprising a first plate, a second plate and a spacer, wherein the spacer is configured to adjust the gap between the plates when the plates compress the sample into a thin layer.

[0847] The apparatus, method or system of any preceding claim, wherein

[0848] The sample holder includes a first plate, a second plate and a spacer, wherein the spacer has a uniform height and a constant spacing distance. and

[0849] The sample is compressed by the sample holder into a thin layer of uniform thickness, which is adjusted by the height of the spacer.

[0850] Apparatus, method or system as claimed in any preceding claim, wherein the sample is compressed into a layer having a uniform thickness substantially equal to the uniform height of the spacers fixed to the one or both plates.

[0851] The apparatus, method or system of any preceding claim, wherein the sample is compressed into a layer of uniform thickness which varies by less than 15%, 10%, 5%, 2%, 1% or either of these two values.

[0852] The apparatus, method or system of any preceding claim, wherein, in the closed configuration, the sample has a thickness of 500 nm or less, 1000 nm or less, 2 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 mm (millimeter) or less, 2 mm or less, 3 mm or less, 5 mm or less, 10 mm or less, or in a range between any two of these values.

[0853] Apparatus, method or system as claimed in any preceding claim, wherein, in the closed configuration, the sample has a thickness in the range 0.5-20 μm.

[0854] The apparatus, method or system of any preceding claim, wherein, in the closed configuration, the gap between the first plate and the second plate is 500 nm or less, 1000 nm or less, 2 μm (micrometers) or less, 5 μm or less, 10 μm or less, 20 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 mm (millimeter) or less, 2 mm or less, 3 mm or less, 5 mm or less, 10 mm or less or a range between any two of these values.

[0855] The apparatus, method or system of any preceding claim, wherein the sample holder comprises a first plate and a second plate, wherein the thickness of each plate is 500 nm or less, 1000 nm or less, 2 μm (micrometers) or less, 5 μm or less, 10 μm or less, 20 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 mm (millimeter) or less, 2 mm or less, 3 mm or less, 5 mm or less, 10 mm, or between any two of these values.

[0856] The apparatus, kit or method of any preceding claim, wherein the imager comprises a camera.

[0857] The device, kit or method of any preceding claim, wherein the imager is part of the detector.

[0858] The apparatus, kit or method of any preceding claim, wherein the imager is integral to the detector.

[0859] The apparatus, kit or method of any preceding claim, wherein the imager is directed by the software to capture one or more images of the sample, identify the interfering element regions and the interfering element-free regions, and digitally distinguish the interfering element regions from the interfering element-free regions.

[0860] The apparatus, kit or method of any preceding claim, wherein the imager comprises a filter configured to filter a signal from the sample.

[0861] The apparatus, kit or method of any preceding claim, wherein the imager comprises a light source configured to illuminate the sample.

[0862] The device, kit or method of any preceding claim, wherein the detector is a mobile device.

[0863] The device, kit or method of any preceding claim, wherein the detector is a smartphone.

[0864] The apparatus, kit or method of any preceding claim, wherein the detector is a smartphone and the imager is a camera that is part of the smartphone.

[0865] The device, kit or method of any preceding claim, wherein the detector comprises a display configured to display the presence and / or amount of the analyte.

[0866] The device, kit or method of any preceding claim, wherein the detector is configured to transmit the detection result to a third party.

[0867] The apparatus, kit or method of any preceding claim, wherein the software is stored in a storage unit which is part of the detector.

[0868] The apparatus, kit or method of any preceding claim, wherein the software is configured to direct the detector to indicate the presence and / or amount of the analyte.

[0869] The apparatus, kit or method of any preceding claim, wherein the software is configured to direct the imager to calculate a combined signal for the analyte from the region free of interfering elements.

[0870] The apparatus, kit or method of any preceding claim, wherein the software is configured to direct the imager to ignore signals from analytes in the region of the interfering element.

[0871] The apparatus, kit or method of any preceding claim, wherein the software is configured to direct the imager to increase signal contrast of signals from areas of the interfering element to signals from areas without the interfering element.

[0872] The apparatus, kit or method of any preceding claim, wherein the software is configured to direct the detector to calculate a ratio of signals from the region of the interfering element to the region of the non-interfering element.

[0873] The apparatus, method or system of any preceding claim, wherein the mobile device is a smartphone.

[0874] The device, method or system of any preceding claim, wherein the mobile device comprises a set of instructions which, when executed, direct the device to capture one or more images of a sample,

[0875] The apparatus, method or system of any preceding claim, wherein the mobile device comprises a light source configured to illuminate the sample.

[0876] The device, method or system of any preceding claim, wherein the mobile device comprises a display configured to display the presence and / or amount of the analyte.

[0877] The device, method or system of any preceding claim, wherein the mobile device comprises a set of instructions which, when executed, direct the detector to display the presence and / or amount of an analyte.

[0878] The apparatus, method or system of any preceding claim, wherein the mobile device is configured to transmit the detection result to a third party.

[0879] The apparatus, method or system of any preceding claim, wherein the adapter comprises a filter configured to filter a signal from the sample.

[0880] The device, method or system of any preceding claim, wherein the adapter includes a slot into which the device can be inserted.

[0881] The device, method or system of any preceding claim, wherein the adapter includes a slide to facilitate insertion of the device into the slot.

[0882] The device, method or system of any preceding claim, wherein the adapter comprises a cradle frame configured to be removably connected to the mobile device.

[0883] The apparatus, method or system of any preceding claim, wherein the adapter comprises an optical cartridge including one or more optical components configured to enhance a signal from the sample.

[0884] The device, method or system of any preceding claim, wherein the device or method is used to detect proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles and other molecules, compounds, mixtures and substances thereof.

[0885] The device, method or system of any preceding claim, wherein the device or method is for use in diagnosing, managing and / or preventing human diseases and conditions.

[0886] The device, method or system of any preceding claim, wherein the device or method is for use in diagnosing, managing and / or preventing veterinary diseases and conditions, or for use in diagnosing, managing and / or preventing plant diseases and conditions.

[0887] The apparatus, method or system of any preceding claim, wherein the apparatus or method is used for environmental testing and decontamination.

[0888] The apparatus, method or system of any preceding claim, wherein the apparatus or method is for use in agricultural or veterinary applications.

[0889] The apparatus, method or system of any preceding claim, wherein the apparatus or method is for use in food testing.

[0890] The device, method or system of any preceding claim, wherein the device or method is used for drug testing and prevention.

[0891] The apparatus, method or system of any preceding claim, wherein the apparatus or method is for detecting and / or measuring an analyte in blood.

[0892] The apparatus, method or system of any preceding claim, wherein the apparatus or method is for use in a colorimetric assay.

[0893] The apparatus, method or system of any preceding claim, wherein the apparatus or method is for use in fluorescence assays.

[0894] Apparatus, method or system as claimed in any preceding claim, wherein the signal associated with the analyte is an electrical signal or an optical signal.

[0895] The apparatus, method or system of any preceding claim, wherein the signal associated with the analyte is an optical signal that allows the imager to capture images of regions enriched in the interfering element and regions depleted in the interfering element.

[0896] The apparatus, method or system of any preceding claim, wherein the signal associated with the analyte is derived from a colorimetric reaction.

[0897] Apparatus, method or system as claimed in any preceding claim, wherein a signal associated with the analyte is generated by illuminating the sample with an illumination source.

[0898] Apparatus, method or system as claimed in any preceding claim, wherein the plates are movable relative to each other.

[0899] The apparatus, method or system of any preceding claim, wherein the spacers are secured to one or both of the plates and are of uniform height.

[0900] The apparatus, method or system of any preceding claim, wherein the first and second plates are configured to compress the sample into a layer of uniform thickness, the thickness being substantially equal to the height of the spacer.

[0901] The apparatus, method or system of any preceding claim, wherein the spacer has a uniform height of 1 mm or less, 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less, 0.1 μm or less, 50 nm or less, 20 nm or less, 10 nm or less, or a range between two values.

[0902] The apparatus, method or system of any preceding claim, wherein the spacers have a uniform height in the range of 0.5-2 μm, 0.5-3 μm, 0.5-5 μm, 0.5-10 μm, 0.5-20 μm, 0.5-30 μm or 0.5-50 μm.

[0903] The apparatus, method or system of any preceding claim, wherein at least one of the plates has a thickness of 100 mm or less, 50 mm or less, 25 mm or less, 10 mm or less, 5 mm or less, 1 mm or less, 500 μm or less, 400 μm or less, 300 μm or less, 200 μm or less, 175 μm or less, 150 μm or less, 125 μm or less, 100 μm or less, 75 μm or less, 50 μm or less, 40 μm or less, 30 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 4 μm or less, 3 μm or less, 2 μm or less, 1.8 μm or less, 1.5 μm or less, 1 μm or less, 0.5 μm or less, 0.2 μm or less or 0.1 μm or less, or a thickness between any two values.

[0904] The apparatus, method or system of any preceding claim, wherein at least one of the plates has a thickness in the range of 0.5 to 1.5 mm; around 1 mm; in the range of 0.15 to 0.2 mm; or approximately 0.175 mm.

[0905] The apparatus, method or system of any preceding claim, wherein at least one of the plates has a thickness of 1 mm 2 Below, 10mm 2 Below, 25mm 2 Below 50mm 2 Below, 75mm 2 Below, 1cm 2 (square centimeters) or less, 2cm 2 Below, 3cm 2 Below, 4cm 2 Below, 5cm 2 Below, 10cm 2 Below 100cm 2 Below 500cm 2 Below 1000cm 2 Below 5000cm 2 Below 10,000cm 2 Below 10,000cm 2 The lateral area below or in the range between two values.

[0906] The apparatus, method or system of any preceding claim, wherein at least one of the plates has a lateral area of ​​500 to 1000 mm 2 and the lateral area of ​​the plate is between 500mm and 1000mm 2 Within the range or about 750mm 2 .

[0907] The apparatus, method or system of any preceding claim, wherein the Young's modulus of the gasket multiplied by the packing factor of the gasket is equal to or greater than 10 MPa, wherein the packing factor is the ratio of the gasket area in contact with the uniform thickness layer to the total plate area in contact with the uniform thickness layer.

[0908] The apparatus, method or system of any preceding claim, wherein the thickness of the flexible sheet multiplied by the Young's modulus of the flexible sheet is in the range of 60 to 750 GPa-μm.

[0909] The apparatus, method or system of any preceding claim, wherein, for a flexible board, the fourth power of the inter-pad distance (ISD) divided by the thickness (h) of the flexible board and the Young's modulus (E) of the flexible board, ISD 4 / (hE) is equal to or less than 10 6 μm 3 / GPa.

[0910] The apparatus, method or system of any preceding claim, wherein one or both plates include position markings on or within the surface of the plate, the position markings providing position information of the plate.

[0911] Apparatus, method or system as claimed in any preceding claim, wherein one or both plates include a scale on the surface or in the interior of the plate which provides information on the lateral dimensions of the structure of the sample and / or the plate.

[0912] Apparatus, method or system as claimed in any preceding claim, wherein one or both plates include image markings on the surface or in the interior of said plates, said image markings facilitating imaging of said sample.

[0913] The apparatus, method or system of any preceding claim, wherein the distance between the spacers is in the range 7 μm to 50 μm.

[0914] The apparatus, method or system of any preceding claim, wherein the distance of the spacers is in the range 50 μm to 120 μm.

[0915] The apparatus, method or system of any preceding claim, wherein the distance of the spacers is in the range 120 μm to 200 μm.

[0916] The apparatus, method or system of any preceding claim, wherein the spacer is a post having a cross-sectional shape selected from the group consisting of circular, polygonal, circular, square, rectangular, elliptical, oval or any combination thereof.

[0917] The apparatus, method or system of any preceding claim, wherein the spacers have a cylindrical shape and have a substantially flat top surface, wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.

[0918] The apparatus, method or system of any preceding claim, wherein each spacer has a ratio of its lateral dimension to its height of at least 1.

[0919] The apparatus, method or system of any preceding claim, wherein the smallest lateral dimension of the spacer is less than or substantially equal to the smallest dimension of the analyte in the sample.

[0920] The apparatus, method or system of any preceding claim, wherein the smallest lateral dimension of the spacer is in the range 0.5 μm to 100 μm.

[0921] The apparatus, method or system of any preceding claim, wherein the smallest lateral dimension of the spacer is in the range of 0.5 μm to 10 μm.

[0922] The apparatus, method or system of any preceding claim, wherein the spacer has a cylindrical shape and the sidewall corners of the spacer are rounded with a radius of curvature of at least 1 μm.

[0923] The apparatus, method or system of any preceding claim, wherein the spacer has a thickness of at least 100 / mm 2 density.

[0924] The apparatus, method or system of any preceding claim, wherein the spacer has a thickness of at least 1000 / mm 2 density.

[0925] The apparatus, method or system of any preceding claim, wherein at least one of the plates is transparent

[0926] The apparatus, method or system of any preceding claim, wherein at least one of the plates is made from a flexible polymer.

[0927] The apparatus, method or system of any preceding claim wherein the gasket is incompressible with respect to pressure compressing the plates and / or independently only one of the plates is flexible.

[0928] The apparatus, method or system of any preceding claim, wherein the flexible sheet has a thickness in the range 10 μm to 200 μm.

[0929] The apparatus, method or system of any preceding claim, wherein the variation in sample thickness is less than 30%.

[0930] The apparatus, method or system of any preceding claim, wherein the variation in sample thickness is less than 10%.

[0931] The apparatus, method or system of any preceding claim, wherein the variation in sample thickness is less than 5%.

[0932] The apparatus, method or system of any preceding claim, wherein the first and second panels are connected and configured to change from an open configuration to a closed configuration by folding the panels.

[0933] The apparatus, method or system of any preceding claim, wherein the first and second panels are connected by a hinge and are configured to change from an open configuration to a closed configuration by folding the panels along the hinge.

[0934] The apparatus, method or system of any preceding claim, wherein the first and second panels are connected to the panels by hinges as separate materials and are configured to change from an open configuration to a closed configuration by folding along the hinges of the panels.

[0935] The apparatus, method or system of any preceding claim, wherein the first and second panels are made from a single piece of material and are configured to change from an open configuration to a closed configuration by folding the panels.

[0936] The apparatus, method or system of any preceding claim, wherein the sample layer of uniform thickness is at least 1 mm 2 It is uniform over the lateral area.

[0937] Apparatus, method or system as claimed in any preceding claim, wherein the shims are secured to the plate by rolling directly onto the plate or by injection moulding the plate.

[0938] The apparatus, method or system of any preceding claim, wherein the material of the plates and the spacers is selected from polystyrene, PMMA, PC, COC, COP or another plastic.

[0939] (1) Definition

[0940] The terms used to describe the apparatus / devices, systems and methods disclosed herein are defined in PCT Applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456,287, filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456,504, filed on February 8, 2017, the entire contents of which are incorporated herein in their entirety for all purposes.

[0941] (2) Sample

[0942] The devices / apparatus, systems and methods disclosed herein can be applied to the manipulation and detection of various types of samples. These samples are disclosed, listed, described and / or summarized in PCT applications PCT / US2016 / 045437 and PCT / US0216 / 051775 filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065 filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287 filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0943] The devices, apparatuses, systems and methods disclosed herein can be used for samples, such as but not limited to diagnostic samples, clinical samples, environmental samples and food samples. The types of samples include but are not limited to those described in PCT applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and the entire contents of which are incorporated herein by reference.

[0944] For example, in some embodiments, the devices, apparatus, systems and methods disclosed herein are used for samples comprising cells, tissues, body fluids and / or mixtures thereof. In some embodiments, the sample comprises a human body fluid. In some embodiments, the sample comprises cells, tissues, body fluids, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, uterus, chyle, chyme, endolymph, peripheral lymph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, mucus, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine and at least one of exhaled breath condensate.

[0945] In some embodiments, the apparatus, devices, systems and methods disclosed herein are used for environmental samples obtained from any suitable source, such as, but not limited to: rivers, lakes, ponds, oceans, glaciers, icebergs, rainwater, snow, sewage, reservoirs, tap water, drinking water, etc.; solid samples from soil, compost, sand, rocks, concrete, wood, bricks, sewage, etc.; in some embodiments, environmental samples are freshly obtained from the source; and gaseous samples from air, underwater vents, industrial exhaust, vehicle exhaust, etc. In some embodiments, the environmental sample is processed. For example, the sample in non-liquid form is converted into liquid form before applying the apparatus, devices, systems and methods of the present invention.

[0946] In some embodiments, the devices, apparatuses, systems and methods disclosed herein are used for food samples that are suitable or have the potential to become suitable for animal consumption, such as human consumption. In some embodiments, food samples include raw materials, cooked or processed foods, plant and animal food sources, pre-processed foods, and partially or completely processed foods, etc. In certain embodiments, samples in non-liquid form are converted to liquid form prior to application of the devices, apparatuses, systems and methods of the present invention.

[0947] The present devices, apparatus, systems and methods can be used to analyze samples of any volume. Examples of volumes include, but are not limited to, about 10 mL or less, 5 mL or less, 3 mL or less, 1 microliter (μL, "uL" herein) or less, 500 μL or less, 300 μL or less, 250 μL or less, 200 μL or less, 170 μL or less, 150 μL or less, 125 μL or less, 100 μL or less, 75 μL or less, 50 μL or less, 25 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, 1 pL or less, or a range between two values.

[0948] In some embodiments, the volume of the sample includes, but is not limited to, about 100 μL or less, 75 μL or less, 50 μL or less, 25 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, 1 pL or less, or a range between two values. In some embodiments, the volume of the sample includes, but is not limited to, about 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, or a range between two values.

[0949] In some embodiments, the amount of the sample is about one drop of liquid. In some embodiments, the amount of the sample is the amount collected from a punctured finger or fingertip. In some embodiments, the amount of the sample is the amount collected from a microneedle, micropipette, or intravenous aspiration.

[0950] In certain embodiments, the sample holder is configured to accommodate a fluid sample. In certain embodiments, the sample holder is configured to compress at least a portion of the fluid sample into a thin layer. In certain embodiments, the sample holder includes a structure configured to heat and / or cool the sample. In certain embodiments, the heating source provides electromagnetic waves, which can be absorbed by certain structures in the sample holder to change the temperature of the sample. In certain embodiments, the signal sensor is configured to detect and / or measure a signal from the sample. In certain embodiments, the signal sensor is configured to detect and / or measure an analyte in the sample. In certain embodiments, the radiator is configured to absorb heat from the sample holder and / or a heat source. In certain embodiments, the radiator includes a chamber that at least partially surrounds the sample holder.

[0951] (3) Q-cards, spacers, and uniform sample thickness

[0952] The apparatus / devices, systems and methods disclosed herein may include or use Q cards, shims and uniform sample thickness embodiments for sample detection, analysis and quantification. In some embodiments, the Q card includes a shim that helps to render at least a portion of the sample as a highly uniform layer. The structure, materials, functions, variations and dimensions of the shim, as well as the uniformity of the shim and sample layer, are disclosed, listed, described and summarized in PCT applications (designated US) PCT / US2016 / 045437 and PCT / US0216 / 051775 filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065 filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287 filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0953] When using the QMAX card, both plates need to be opened first for sample deposition. However, in some embodiments, the QMAX card from the package has two plates in contact with each other (e.g., closed position), and it is difficult to separate them because one or both plates are very important. To facilitate the opening of the QMAX card, one or more opening notches are created at the edge or corner of the first plate or both positions, and in the closed position of the plate, a portion of the second plate is placed on the opening notches, so that the second plate can be lifted up in the notch of the first plate without blocking the first plate.

[0954] In the QMAX analysis platform, the QMAX card uses two plates to control the shape of the sample into a thin layer (e.g., by compression). In some embodiments, the manipulation of the plates requires several changes in the relative position of the two plates (referred to as: plate configuration) by human hands or other external forces. The QMAX card needs to be designed to make manual operation simple and fast.

[0955] In the QMAX analysis, one of the plate configurations is an open configuration, in which the two plates are completely or partially separated (the spacing between the plates is not controlled by the spacer), and the sample can be deposited. The other configuration is a closed configuration, in which at least a portion of the sample deposited in the open configuration is compressed by the two plates into a layer of highly uniform thickness, the uniform thickness of the layer being limited by the inner surface of the plates and regulated by the plates and spacers. In some embodiments, the average spacing between the two plates is greater than 300 μm.

[0956] In the operation of the QMAX analysis, the operator first needs to place both plates in an open state in preparation for sample deposition, then deposit the sample on one or both plates, and finally close the plates to a closed position. In certain embodiments, the two plates of the QMAX card are initially stacked on top of each other and need to be separated to enter the open configuration for sample deposition. When one of the plates is a plastic film (175 micron thick PMA), this separation is difficult to perform by hand. The present invention is intended to provide an apparatus and method that makes the operation of certain assays, such as QMAX card assays, easy and rapid.

[0957] In some embodiments, the QMAX device includes a hinge that connects two or more panels together so that the panels can be opened and closed in a manner similar to a book. In some embodiments, the material of the hinge is such that after adjustment, the hinge can self-maintain the angle between the panels. In some embodiments, the hinge is configured to hold the QMAX card in a closed configuration so that the entire QMAX card can slide in and out of the card slot without causing accidental separation of the two panels. In some embodiments, the QMAX device includes one or more hinges that can control the rotation of more than two panels.

[0958] In some embodiments, the hinge is made of a metal material selected from gold, silver, copper, aluminum, iron, tin, platinum, nickel, cobalt, alloys or any combination thereof. In some embodiments, the hinge comprises a single layer made of a polymer material, such as but not limited to plastic. The polymer material is selected from acrylate polymers, vinyl polymers, olefin polymers, cellulosic polymers, non-cellulosic polymers, polyester polymers, nylon, cyclic olefin copolymers (COC), poly(methyl methacrylate) (PMMB), polycarbonate (PC), cyclic olefin polymers (COP), liquid crystal polymers (LCP), polyamide (PB), polyethylene (PE), polyimide (PI), polypropylene (PP), polyphenylene ether (PPE), polystyrene (PS), polyoxymethylene (POM), polyetheretherketone (PEEK), polyethersulfone (PES), polyethylene phthalate (PET), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), polyvinylidene fluoride (PVDF), polybutylene terephthalate (PBT), fluorinated ethylene propylene (FEP), perfluoroalkoxyalkane (PFB), polydimethylsiloxane (PDMS), rubber or any combination thereof. In some embodiments, the polymer material is selected from polystyrene, PMMB, PC, COC, COP, other plastics, or any combination thereof.

[0959] Essentially, unless otherwise specified, the term "shim" or "stopper" may refer to a mechanical object that is set to limit the minimum spacing between two plates that can be achieved when the two plates are compressed. That is, in compression, the shim will stop the relative movement of the two plates to prevent the plate spacing from becoming less than a preset (i.e., predetermined) value.

[0960] In some embodiments, a person's hand can be used to press the plate into a closed state; in some embodiments, a person's hand can be used to press the sample into a thin layer. The manner of using hand pressure is described and / or summarized in PCT applications (designating the United States) PCT / US2016 / 045437 filed on August 10, 2016 and PCT / US0216 / 051775 filed on September 14, 2016, Provisional Application No. 62 / 431,639 filed on December 9, 2016, No. 62 / 456,287 filed on February 8, 2017, No. 62 / 456,065 filed on February 7, 2017, No. 62 / 456,504 filed on February 8, 2017, and Provisional Application No. 62 / 460,062 filed on February 16, 2017. The entire contents of which are incorporated herein by reference.

[0961] In some embodiments, a human hand can be used to manipulate or manipulate the plates of the QMAX device. In some embodiments, a human hand can be used to apply an imprecise force to compress the plates from an open configuration to a closed configuration. In some embodiments, a human hand can be used to apply an imprecise force to achieve a high degree of uniformity in sample thickness (e.g., less than 5%, 10%, 15%, or 20% variation).

[0962] (4) Hinges, slots, concave edges and slides

[0963] The devices / apparatuses, systems and methods disclosed herein may include or use Q-cards for sample detection, analysis and quantification. In some embodiments, the Q-cards include hinges, notches, grooves and slides, which help facilitate the manipulation of the Q-cards and the measurement of samples. The structures, materials, functions, variations and dimensions of the hinges, notches, recesses and slides disclosed herein are disclosed, listed, described and / or summarized in PCT applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application 62 / 431,639, filed on December 9, 2016, U.S. Provisional Application 62 / 256,065, filed on February 7, 2017, U.S. Provisional Application 62 / 456,287 and 62 / 456,504, filed on February 8, 2017, and U.S. Provisional Application 62 / 539,660, filed on August 1, 2017, which are incorporated herein in their entirety for all purposes.

[0964] In some embodiments, the QMAX device includes an opening mechanism, such as but not limited to a notch on the edge of the board or a strap attached to the board, making it easier for the user to manipulate the positioning of the board, such as but not limited to separating the boards by hand.

[0965] In some embodiments, the QMAX device includes grooves on one or both plates. In certain embodiments, the grooves restrict the flow of the sample on the plates.

[0966] (5) Q card and adapter

[0967] The devices / apparatus, systems and methods disclosed herein may include or use a Q card for sample detection, analysis and quantification. In some embodiments, the Q card is used with an adapter configured to accommodate the Q card and connect to a mobile device so that the sample in the Q card can be imaged, analyzed and / or measured. Mobile device. The structure, materials, functions, variations, dimensions and connections of the Q card, adapter and mobile phone are disclosed, listed, described and / or summarized in PCT applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775 filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065 filed on February 7, U.S. Provisional Application Nos. 62 / 456287 and 62 / 456590 filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017 459,544, filed February 15, 2017, and U.S. Provisional Applications 62 / 460075 and 62 / 459920, filed February 16, 2017, are hereby incorporated in their entirety for all purposes.

[0968] In some embodiments, the adapter includes a socket slot configured to accommodate the QMAX device when the device is in a closed configuration. In some embodiments, the QMAX device has a sample deposited therein, and the adapter can be connected to a mobile device (e.g., a smartphone) so that the sample can be read by the mobile device. In some embodiments, the mobile device can detect and / or analyze signals from the sample. In some embodiments, the mobile device can capture an image of the sample while the sample is in the QMAX device and in the field of view (FOV) of a camera, in some embodiments, the camera is part of the mobile device.

[0969] In some embodiments, the adapter includes an optical assembly configured to enhance, amplify and / or optimize the generation of a signal from the sample. In some embodiments, the optical assembly includes a component configured to enhance, amplify and / or optimize the illumination provided to the sample. In certain embodiments, the illumination is provided by a light source that is part of the mobile device. In some embodiments, the optical assembly includes a component configured to enhance, amplify and / or optimize the signal from the sample.

[0970] (6) Smartphone detection system

[0971] The devices / apparatus, systems and methods disclosed herein may include or use a Q card for sample detection, analysis and quantification. In some embodiments, the Q card is used with an adapter that can connect the Q card to a smartphone detection system. In some embodiments, the smartphone includes a camera and / or an illumination source. The smartphone detection system and associated hardware and software herein are disclosed, listed, described and / or summarized in PCT applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application Nos. 62 / 456287 and 62 / 456590, filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, U.S. Provisional Application No. 459,544, filed on February 15, 2017, and U.S. Provisional Application Nos. 62 / 460075 and 62 / 459920, filed on February 16, 2017, all of which are incorporated herein in their entirety for all purposes.

[0972] In some embodiments, the smartphone includes a camera that can be used to capture images or samples when the sample is placed in the camera's field of view (e.g., via an adapter). In some embodiments, the camera includes a set of lenses (e.g., in an iPhone TM 6). In some embodiments, the camera includes at least two sets of lenses (e.g., in an iPhone TM 7). In some embodiments, the smartphone includes a camera, but the camera is not used for image capture.

[0973] In some embodiments, the smartphone includes a light source such as, but not limited to, an LED (light emitting diode). In some embodiments, the light source is used to provide illumination to the sample when the sample is in the field of view of the camera (e.g., through an adapter). In some embodiments, the light from the light source is enhanced, amplified, altered, and / or optimized by optical components of the adapter.

[0974] In some embodiments, the smartphone includes a processor configured to process information from the sample. The smartphone includes software instructions that, when executed by the processor, can enhance, amplify and / or optimize the signal (e.g., image) from the sample. The processor may include one or more hardware components, such as a central processing unit (CPU), an application specific integrated circuit (ASIC), an application specific instruction set processor (ASIP), a graphics processing unit (GPU), a physical processing unit (PPU), a digital signal processor (DSP), a field programmable gate array (FPGA), a programmable logic device (PLD), a controller, a microcontroller unit, a reduced instruction set computer (RISC), a microprocessor, etc., or any combination thereof.

[0975] In some embodiments, the smartphone includes a communication unit configured and / or used to transmit data and / or images related to the sample to another device. By way of example only, the communication unit can use a cable network, a wired network, an optical fiber network, a telecommunications network, an intranet, the Internet, a local area network (LAN), a wide area network (WAN), a wireless local area network (WLAN), a metropolitan area network (MAN), a wide area network (WAN), a public switched telephone network (PSTN), a Bluetooth network, a ZigBee network, a near field communication (NFC) network, or the like, or any combination thereof.

[0976] In some embodiments, the smartphone is an iPhone TM , Android TM Phone or Windows TM Telephone.

[0977] (7) Detection method

[0978] The devices / apparatus, systems and methods disclosed herein may include or be used for various types of detection methods. The detection methods disclosed herein are disclosed, listed, described and / or summarized in PCT applications PCT / US2016 / 045437 and PCT / US0216 / 051775 filed on August 10, 2016 and September 14, 2016, U.S. Provisional Application No. 62 / 456065 filed on February 7, 2017, U.S. Provisional Application Nos. 62 / 456287, 62 / 456528, 62 / 456631, 62 / 456522, 62 / 456598, 62 / 456603 and 62 / 4 56628, filed on February 8, 2017, U.S. Provisional Applications 62 / 459276, 62 / 456904, 62 / 457075 and 62 / 457009, filed on February 9, 2017, and U.S. Provisional Applications 62 / 459303, 62 / 459337 and 62 / 459598, filed on February 15, 2017 and U.S. Provisional Applications 62 / 460083, 62 / 460076, filed on February 16, 2017, all of which are incorporated herein in their entirety for all purposes.

[0979] (8) Labels, capture agents and detection agents

[0980] The devices / apparatus, systems and methods disclosed herein may employ various types of labels, capture agents and detectors for analyte detection. The labels herein are disclosed, listed, described and / or summarized in PCT applications (designated US) PCT / US2016 / 045437 and PCT / US0216 / 051775 filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065 filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287 filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504 filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0981] In some embodiments, the label is optically detectable, such as, but not limited to, a fluorescent label. In some embodiments, labels include, but are not limited to, IRDye800CW, Alexa 790, Dylight 800, fluorescein, fluorescein isothiocyanate, succinimidyl ester of carboxyfluorescein, succinimidyl ester of fluorescein, 5-isomer of fluorescein dichlorotriazine-alanine-box carboxyalanine, Oregon Green 488, Oregon Green 514; Lucifer Yellow, alpha-pyridinium orange, rhodamine, tetramethylrhodamine, Texas Red, propidium iodide, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide), tetrabromo-rhodamine 123, rhodamine 6G, TMRM (tetramethylrhodamine methyl ester) , TMRE (tetramethylrhodamine ethyl ester), tetramethylrhodamine, rhodamine B and 4-dimethylaminotetramethylrhodamine, green fluorescent protein, blue-shifted green fluorescent protein, blue-shifted green fluorescent protein, red-shifted green fluorescent protein, yellow-shifted green fluorescent protein, 4-acetylamino-4'-isothiocyanatostyrene-2,2'disulfonic acid; cridine and its derivatives, such as a-cridine, a-cridine isothiocyanate; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS); 4-amino-N-[3-vinylsulfonyl)phenyl]naphthamide-3.5-disulfonate; N-(4-anilino-1-naphthyl)maleimide; anthracenamide 4,4-difluoro-5-(2-thienyl)-4-boron-3a,4a-diaza-5-indenotri-3-propionic acid BODIPY; Cascade Blue; Brilliant Yellow; Coumarin and its derivatives: coumarin, 7-amino-4-methylcoumarin (AMC, coumarin 120), 7-amino-4-trifluoromethylcoumarin (coumarin 151); cyanine dyes; anthocyanin 4 ',6-Diaminodiamino-2-phenylindole (DAPI); 5',5"-dibromopyrogallol-sulfonphthalein (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; diethylenetriamine pentaacetate 4,4'-diisothiocyanatodihydro-styrene-2-,2'-disulfonic acid; 4,4'-diisothiocyanatostyrene-2,2'-disulfonic acid; 5-(dimethylamino)naphthalene-1-sulfonyl chloride (DNS, dansyl chloride); 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC); Eosin and its derivatives: Eosin, Eosin isothiocyanate, Erythrosin and its derivatives: Eosin B, Eosin, isothiocyanate; Ethidium; Fluorescein and its derivatives: 5-carboxyfluorescein (FAM), 5-(4,6-dichlorotriazin-2-yl)amino-fluorescein (DTAF), 2',7'-dimethoxy-4'5'-dichloro-6- Carboxyfluorescein (JOE), fluorescein, fluorescein isothiocyanate, QFITC ​​(XRITC); fluorescamine; IR144; IR1446; malachite green isothiocyanate; 4-methylumbelliferyl-ferrophthalene; nitrotyrosine; p-aniline; phenol red; B-phycoerythraldehyde; o-phthalaldehyde; catechol; pyr, pyr butyrate, succinimidyl 1-py; butyrate quantum dots; reactive red 4 (Cibacron. TMBrilliant Red 3B-A) Rhodamine and its derivatives: 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), lysine rhodamine B sulfonyl chloride rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine X isothiocyanate, sulforhodamine B, sulforhodamine 101, sulfonyl chloride derivative of sulforhodamine 101 (Texas Red); N,N,N',N'-tetramethyl-6-carboxyrhodamine (TAMRA); tetramethylrhodamine; tetramethylrhodamine isothiocyanate (TRITC); riboflavin; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS), 4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL), rosmarinic acid; CAL fluorescent orange 560; che chelate derivatives; Cy 3; Cy 5; Cy 5.5; Cy 7; IRD 700; IRD 800; La Jolla Blue Phthalocyanine; and naphthalocyanines, coumarins and related dyes, xanthene dyes such as rhododendrons, resorcinols, dienes, azides, isoindole, dansyl dyes, aminophthalhydrazides (such as luminol) and isoluminol derivatives, aminophthalimides, aminonaphthalimides, aminobenzofurans, aminoquinolines, aminoquinolines, complexes; combinations thereof, and the like. Suitable fluorescent proteins and chromogenic proteins include, but are not limited to, green fluorescent protein (GFP), including, but not limited to, GFP derived from Aequoriavictoria or a derivative thereof, such as a "humanized" derivative such as Enhanced GFP; a GFP from another species, such as Renilla renilla, Renilla renilla, or Ptilosarcus guernyi; a "humanized" recombinant GFP (hrGFP); any of a variety of fluorescent and colored proteins from Anthozoan species; combinations thereof; and the like.

[0982] In any embodiment, the QMAX device may comprise a plurality of capture agents and / or detector agents, each of which binds a biomarker selected from Tables B1, B2, B3 and / or B7 in U.S. Provisional Application No. 62 / 234,538 and / or PCT / US2016 / 054025. Wherein the reading step d) comprises obtaining a measure of the amount of the plurality of biomarkers in the sample, and wherein the amount of the plurality of biomarkers in the sample is used for diagnosis of a disease.

[0983] In any embodiment, the capture agent and / or the detection agent can be an antibody antigenic determinant, and the biomarker can be an antibody that binds to the antibody antigenic determinant. In some embodiments, the antibody antigenic determinant includes a biomolecule or fragment thereof selected from Table B4, B5 or B6 in U.S. Provisional Application No. 62 / 234,538 and / or PCT Application No. PCT / US2016 / 054025. In some embodiments, the antibody antigenic determinant includes an allergen or fragment thereof selected from Table B5. In some embodiments, the antibody antigenic determinant includes a biomolecule or fragment thereof derived from an infectious agent selected from Table B6 in U.S. Provisional Application No. 62 / 234,538 and / or PCT Application No. PCT / US2016 / 054025.

[0984] In any embodiment, the QMAX device may comprise a plurality of antibody antigenic determinants selected from Tables B4, B5 and / or B6 of U.S. Provisional Application No. 62 / 234,538 and / or PCT Application No. PCT / US2016 / 054025, wherein the reading step d) comprises obtaining a measure of the amount of the plurality of antigenic determinant binding antibodies in the sample, and wherein the amount of the plurality of antigenic determinant binding antibodies in the sample can be used for diagnosis of a disease or condition.

[0985] (9) Analyte

[0986] The devices / apparatus, systems and methods disclosed herein can be applied to the manipulation and detection of various types of analytes, including biomarkers. The analytes disclosed herein are disclosed, listed, described and / or summarized in PCT applications (designating the United States) PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, all of which are hereby incorporated in their entirety for all purposes.

[0987] The devices, apparatuses, systems and methods disclosed herein can be used for the detection, purification and / or quantification of various analytes. In some embodiments, the analytes are biomarkers associated with various diseases. In some embodiments, the analytes and / or biomarkers indicate the presence, severity and / or stage of a disease. Analytes, biomarkers and / or diseases that can be detected and / or measured using the devices, apparatuses, systems and / or methods of the present invention are listed, described and / or summarized in PCT application (designating the United States) PCT / US2016 / 045437 filed on August 10, 2016, PCT application PCT / US2016 / 054025 filed on September 27, 2016, and U.S. Provisional Application Nos. 62 / 234,538 filed on September 29, 2016, 62 / 233,885 filed on September 28, 2015, 62 / 293,188 filed on February 9, 2016, and 62 / 305,123 filed on March 8, 2016, the entire contents of which are incorporated herein by reference. For example, the devices, apparatuses, systems and methods disclosed herein can be used for (a) detection, purification and quantification of compounds or biomolecules associated with the stages of certain diseases (e.g., infectious and parasitic diseases, injuries, diseases, etc.). cardiovascular disease, cancer, psychiatric disease, neuropsychiatric disease and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms (such as viruses, fungi and bacteria) in the environment (such as water, soil) or biological samples (such as tissues, body fluids), (c) detection, quantification of compounds or biological samples that pose a risk to food safety or national security (such as toxic waste, anthrax, etc.),...

Claims

1. A device comprising: The first board, the second board, the light guide spacer (LGS), the sampling area and the reference area, where: (i) the first plate and the second plate are configured to sandwich a sample in a thin layer between the plates for light transmission analysis, and each plate has a sample contact area on its inner surface for contact with the sample; (ii) a light guide spacer (LGS) having a cylindrical shape and sandwiched between the two plates, with both ends of the column directly contacting each plate to form an LGS-plate contact area, thereby forming a structure allowing light to be transmitted from the first plate through the LGS to the second plate without passing through the sample, (iii) a sampling area is an area where light can pass through the first plate, the sample, and the second plate in sequence, wherein the sampling area does not have an LGS; and (iv) the reference area is the area where light passes through the first plate, the light guide spacer, and the second plate in sequence without passing through the sample; Among them, the LGS contact surface and the cross-sectional scale of LGS are larger than the wavelength of light. wherein the light guide spacer is surrounded by the sample or is adjacent to the sample; and Here, the thickness of the sample in the sampling area is 500 μm or less.

2. A device for analyzing hemoglobin in a sample, comprising: A first board, a second board and a light guide spacer, wherein: (i) the first panel and the second panel are movable relative to each other to form different configurations, including an open configuration and a closed configuration; (ii) each plate comprises an inner surface having a sample contact area for contacting a sample containing or potentially containing hemoglobin; and (iii) the light-guiding gasket has a columnar shape, wherein the top surface and the bottom surface of the light-guiding gasket are substantially flat, and the bottom surface is fixed to the inner surface of one of the plates, wherein the bottom surface and the average cross-section of each gasket are respectively larger than the wavelength of light of the analyzed sample, and wherein the light-guiding gasket is within the sample contact area. Among them, an open configuration is a configuration in which: the two plates are separated, the spacing between the plates is not limited by the light-guiding spacer, and the sample is deposited on one or both plates. wherein the closed configuration is the configuration of the sample after deposition in the open configuration; in the closed state: at least a portion of the sample is compressed by the two plates into a highly uniform layer, wherein the uniform thickness of the layer is limited by the plates and the light guide spacers in the sample area of ​​the plates; and wherein, in a closed configuration: (a) at least one gasket in the sample contact area has its top surface in direct contact with one of the plates, and at least one gasket and areas on the plates above and below it define a reference area, wherein the reference area is transparent to light within a specific wavelength range, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate are not occupied by a light-guiding gasket, thereby defining a sampling area that is transparent to light in the same specific wavelength range.

3. A device for analyzing hemoglobin in an analyte in a sample, comprising: A first board, a second board and a light guide spacer, wherein: (i) the first plate and the second plate are configured to receive a sample that contains or may contain an analyte, wherein at least a portion of the sample is between and in contact with the two plates; and (ii) the light guide spacer has a columnar shape and a predetermined basic height, wherein the top and bottom surfaces of the light-guiding spacers are substantially flat, and the top and bottom surfaces of at least one spacer are in direct contact with the board, wherein the top and bottom surfaces of each spacer and the average cross-section thereof are larger than the wavelength of light used to analyze the sample, and wherein: (a) at least one spacer and areas of the plates above and below it define a reference area that is transparent to light within a specific wavelength range and through which light can pass through the plates and spacers, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate is not occupied by a light-guiding spacer, thereby defining a sampling area that is transparent to light within the same wavelength range.

4. A device for analyzing an analyte in a sample, comprising: A first board, a second board and a light guide spacer, wherein: (i) the first panel and the second panel are movable relative to each other into different configurations, including an open configuration and a closed configuration; (ii) each plate comprises an inner surface having a sample contact area for contacting a sample that contains or may contain an analyte; and (iii) the light-guiding spacers have a cylindrical shape and have a predetermined substantially uniform height, wherein the top and bottom surfaces of the light-guiding spacers are substantially flat and the bottom surface is secured to the inner surface of the one or more plates, wherein the top and bottom surfaces and the average cross-sectional dimension of each spacer are respectively larger than the wavelength of light for analyzing the sample, wherein at least one of the light-guiding spacers is within the sample contact region; Among them, an open configuration is a configuration in which: the two plates are separated, the spacing between the plates is not limited by the light-guiding spacer, and the sample is deposited on one or both plates. wherein the closed configuration is the configuration formed after the sample is deposited in the open configuration; in the closed state: at least a portion of the sample is compressed by the two plates into a highly uniform layer, wherein the uniform thickness of the layer is limited by the plates and the light guide spacer in the sample area of ​​the plates; and wherein, in a closed configuration, (a) at least one gasket in the sample contact area has its top surface in direct contact with one of the plates, and at least one gasket and the plate areas above and below it define a reference area, wherein the reference area is transparent to light within a specific wavelength range, and (b) at least one area in the sample contact area on one plate and its corresponding area on the other plate are not occupied by a light-guiding gasket, thereby defining a sampling area that is transparent to light within a specific wavelength range.

5. A device for analyzing hemoglobin in a sample, comprising: A first board, a second board and a light guide spacer, wherein: (v) the first plate and the second plate are configured to sandwich the sample in the thin layer; A first board, a second board and a light guide spacer, wherein: (i) the first plate and the second plate are configured to sandwich the sample in the thin layer; (ii) the light-guiding spacer has a cylindrical shape and is sandwiched between two plates, wherein the head end and the tail end of the column directly contact one of the plates, so that there is no sample between the two ends of the column and the corresponding plate, wherein the light-guiding spacer is surrounded by the sample or in the vicinity of the sample, wherein the direct contact area and the average cross-section of the column are at least 1 um^2 (square micrometer) or greater, respectively; The interval between the inner surfaces of the two plates is 200 μm or less.

6. A sample analysis device, comprising: Any one of the devices in the existing device embodiments, a light source, a camera; and adapters, where (i) the light source is arranged to emit light within a certain wavelength range, and the light within the wavelength range can pass through the reference area; (ii) The camera is configured to image the reference area and the sampling area. (iii) The adapter is configured to position the device, the light source and the camera relative to each other so that light from the light source passes through the reference region and the sampling region and is imaged by the camera.

7. The apparatus of any preceding apparatus embodiment, further comprising: A processor is configured to process the image captured by the camera and determine a property of an analyte in the sample by comparing light transmission from the reference region and the sampling region.

8. An apparatus according to any preceding claim, wherein: The camera and the processor are part of a single mobile device.

9. An apparatus according to any preceding claim, wherein: The light source and the processor are part of a single mobile device.

10. A method for sample analysis using transmitted light, comprising the following steps: (a) a device having any of the preceding apparatus embodiments; (b) depositing a sample in an open configuration of the device, wherein the sample may contain an analyte; (c) placing the device into a closed configuration; (d) constructing a light source having a certain wavelength to pass through the reference region of the device; (e) constructing an imager for imaging the reference region and the sampling region of the device; (f) constructing an adapter configured to position the device, the light source, and the camera relative to each other so that light from the light source passes through the reference region and the sampling region and is imaged by the camera; (g) Determine the nature of the analyte by comparing the light transmittance from the sample area and the reference area.

Images