Low-cost portable microscope imaging system

Abstract

The present invention provides a microscope imaging apparatus, system, and method. The microscope imaging apparatus includes a lens assembly, a first light source assembly, and one or more detection assemblies. The lens assembly has a relatively short working distance, is relatively lightweight, and / or has multiple optical elements. The first light source assembly is configured to irradiate and / or excite a sample placed at the working distance of the lens assembly. One or more detection assemblies are configured to detect light from the sample. The lens assembly, the first light source assembly, and at least one detection assembly are arranged to form an epitaxial configuration.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority based on U.S. Provisional Patent Application No. 63 / 488,992, filed on March 8, 2023. The disclosure of this application is hereby incorporated by reference in its entirety for all purposes.

[0002] This disclosure relates to an imaging device, a system, and a method. In particular, this disclosure relates to a microscope imaging device that is small, lightweight, and low - cost, a system having the microscope imaging device, and a method of using the microscope imaging device and the system for analyzing a biological sample.

Background Art

[0003] In microscopy, an objective lens is typically designed to provide a long focal length and a wide field of view. A typical microscope lens costs at least several hundred dollars. Also, a typical microscope lens weighs about 50 grams or more, so some operating force is required to move the microscope lens (e.g., to adjust the focus). Components that can provide the required force, such as voice - coil motors, are often large and expensive. In some cases, a voice - coil motor alone can cost thousands of dollars. Therefore, many existing microscopes are expensive and bulky and are used in laboratories. These are generally not suitable for personal use at home, medical field applications, etc., and are not affordable.

Summary of the Invention

[0004] Considering the above - mentioned background art, in this technical field, there is still a need for microscope imaging devices, systems, and methods that are suitable not only for hospitals / clinics but also for use in home and medical field applications and are affordable. This disclosure addresses these and other needs in this technical field by providing an affordable portable microscope imaging device and a suitable system and method for analyzing biological samples such as body fluids.

[0005] In one embodiment, the present disclosure provides an imaging device that is small, lightweight, and inexpensive. The imaging device includes a lens assembly, a first light source assembly, and one or more detection assemblies. The lens assembly includes a composite lens and is configured to communicate optically with a sample chamber in the detection area. The lens assembly has (i) an optical path of less than about 15 mm, (ii) a working distance of about 5 mm or less, (iii) a weight of about 50 grams or less, and (iv) more than three optical elements, or any combination thereof. The first light source assembly includes a first light source and is configured to emit light into the sample chamber in the detection area. The first light source assembly is located in the sample chamber on the same side as the lens assembly. One or more detection assemblies are in optical communication with the sample chamber in the detection area and are configured to detect light from the sample chamber through the lens assembly in the detection area. The imaging device is configured to define a first optical path through which light emitted from the first light source passes through the lens assembly and is guided into the sample chamber in the detection area. The imaging device is also configured to define a second optical path through which light returning from the sample chamber passes through a lens assembly in the detection area on the sample stage assembly, and at least a portion of the light returning from the sample chamber in the detection area is directed to one or more detection assemblies.

[0006] In some embodiments, the first optical element among the three or more optical elements closest to the object-side plane of the lens assembly has a first effective aperture diameter and is at a first distance from the object-side plane of the lens assembly. The second optical element among the three or more optical elements furthest from the object-side plane of the lens assembly has a second effective aperture diameter and is at a second distance from the object-side plane of the lens assembly. The first effective aperture diameter is greater than or equal to the second effective aperture diameter, and the first distance is at least 0.4 times the second distance. In some embodiments, the first distance defines the working distance of the lens assembly.

[0007] In some embodiments, the working distance of the lens assembly is less than approximately 4 mm, less than 3 mm, less than 2 mm, or less than 1.5 mm. In some embodiments, the working distance of the lens assembly is approximately 1.0 mm, approximately 1.2 mm, approximately 1.4 mm, or approximately 1.6 mm. In some embodiments, the dimensions of the imaging device are less than 25 centimeters (cm) in length, less than 25 cm in width, and less than 25 cm in height.

[0008] In some embodiments, the composite lens of the lens assembly has an optical axis and an image plane through which the optical axis passes. The composite lens includes a plastic lens barrel surrounding the optical axis. The plastic lens barrel includes an image-side portion through which the optical axis passes and an object-side aperture. The imaging device is used so that the sample is positioned at a distance expressed in the design as the rear focal length, within the limited focal plane of the composite lens. In some embodiments, the composite lens includes two imaging lenses having two different rear focal lengths. The object side uses the imaging lens with the shorter rear focal length, and the image side uses the imaging lens with the longer rear focal length. The effective magnification of the entire imaging device is determined by the ratio of the rear focal lengths of the two imaging lenses. This configuration allows for the use of various imaging lens designs that can result in magnifications of 1x, 2x, 5x, 10x, 15x, 20x, or greater. In some embodiments, the composite lens includes 2 to 10 lenses. In some embodiments, the composite lens is a general-purpose lens that is reversed and positioned so that the sample chamber is located at the rear focal plane of the general-purpose lens.

[0009] In some embodiments, the first light source assembly is configured to emit light in a single narrow wavelength band. In some embodiments, the full width at half maximum (FWHM) of the single narrow wavelength band is 50 nm or less, 25 nm or less, 10 nm or less, or 5 nm or less. In some embodiments, the first light source assembly includes one or more light-emitting diodes (LEDs). In some embodiments, the one or more LEDs include a first LED configured to emit light in a first narrow wavelength band and a second LED configured to emit light in a second narrow wavelength band. In one embodiment, the first light source is a single LED.

[0010] In some embodiments, the imaging device also includes a first dichroic mirror positioned in a first optical path between the lens assembly and a first light source assembly, and in a second optical path between the lens assembly and one or more detection assemblies. The first dichroic mirror has a passband that includes either (a) an excitation wavelength band emitted from the first light source, or (b) one or more emission wavelength bands. The first dichroic mirror also has a stopband that includes the other of (a) an excitation wavelength band emitted from the first light source, or (b) one or more emission wavelength bands. In one embodiment, the first dichroic mirror transmits excitation light emitted from the first light source assembly toward the lens assembly. The lens assembly directs the excitation light to an area of ​​the sample. The lens assembly focuses the synchroic light emitted from one or more components of the sample and passes the focused synchroic light through the first dichroic mirror. The first dichroic mirror reflects the synchroic light into a second optical path different from the first optical path. At least one of the one or more detection assemblies is in optical communication with a first dichroic mirror and detects the synchroic light reflected by the first dichroic mirror. In an alternative embodiment, the first dichroic mirror reflects excitation light emitted from a first light source assembly to a lens assembly. The lens assembly directs the excitation light to an area of ​​the sample. The lens assembly focuses the synchroic light emitted from one or more components of the sample and passes the focused synchroic light through the first dichroic mirror. The first dichroic mirror transmits the synchroic light toward a second optical path different from the first optical path. At least one of the one or more detection assemblies is in optical communication with the first dichroic mirror and detects the synchroic light that has passed through the first dichroic mirror.

[0011] In some embodiments, the imaging device includes a second dichroic mirror positioned in a second optical path between a first dichroic mirror and one or more detection assemblies. The second dichroic mirror has a passband including a first portion of one or more emission wavelength bands and a stopband including a second portion of one or more emission wavelength bands. One or more detection assemblies include a first detection assembly and a second detection assembly. The first detection assembly includes a first two-dimensional photodetector configured to acquire an image in the wavelength band of a first portion of one or more emission wavelength bands. The second detection assembly includes a second two-dimensional photodetector configured to acquire an image in the wavelength band of a second portion of one or more emission wavelength bands. In some embodiments, the first detection assembly includes a first tube lens positioned between the second dichroic mirror and the first two-dimensional photodetector, and the second detection assembly includes a second tube lens positioned between the second dichroic mirror and the second two-dimensional photodetector.

[0012] In some embodiments, the second dichroic mirror is configured to transmit synchrotron radiation in a first wavelength range and reflect synchrotron radiation in a second wavelength range different from the first wavelength range. One or more detection assemblies include a first detection assembly that detects synchrotron radiation in the first wavelength range and a second detection assembly that detects synchrotron radiation in the second wavelength range.

[0013] Alternatively, in some embodiments, the imaging device includes at least one splitter and a plurality of filters. One or more detection assemblies comprise a plurality of detection assemblies. At least one splitter is positioned in a second optical path between a first dichroic mirror and the plurality of detection assemblies and is configured to divide one or more emission wavelength bands into a plurality of parts. Each filter of the plurality of filters is positioned between at least one splitter and a corresponding detection assembly within the plurality of detection assemblies and has its own passband in a plurality of passbands, filtering the corresponding part within the plurality of parts of one or more emission wavelength bands. The corresponding detection assembly within the plurality of detection assemblies includes a two-dimensional photodetector configured to acquire an image of the corresponding part within the plurality of parts of one or more emission wavelength bands after passing through each filter of the plurality of filters.

[0014] In some embodiments, the imaging device includes a voice coil actuator coupled to or integrated with the lens assembly for adjusting the focus of the composite lens. In some embodiments, the voice coil actuator is smaller than 20 mm in width, 20 mm in length, and 20 mm in height. In some embodiments, the focus of the composite lens in the lens assembly is adjustable within a range of approximately ±1.2 mm, approximately ±1.1 mm, approximately ±1.0 mm, approximately ±0.9 mm, or approximately ±0.8 mm or less.

[0015] In some embodiments, the imaging device includes a second light source assembly positioned opposite the sample stage assembly as a lens assembly. The second light source assembly is configured to emit light into the sample chamber in the detection area. The imaging device is configured to define a third optical path through which the light emitted from the second light source assembly passes into the sample chamber and lens assembly in the detection area, and through which at least a portion of the light that has passed through the sample chamber in the detection area is directed to one or more detection assemblies. In some embodiments, the second light source assembly includes a bright-field light source, a dark-field light source, and / or a side-scatter light source. The lens assembly is positioned to focus at least a portion of the light scattered or transmitted by the sample and to pass the focused light through one or more detection assemblies. One or more detection assemblies are configured to detect the light that has passed through the lens assembly.

[0016] In some embodiments, the control unit is configured to selectively operate a first light source assembly, a second light source assembly, any individual light source within the first light source assembly, any individual light source within the second light source assembly, or any combination thereof.

[0017] In some embodiments, the imaging device includes a sample stage assembly configured to position the sample chamber in the detection area. For example, in some embodiments, the sample stage assembly is configured to move the sample chamber relative to the lens assembly. In some embodiments, the sample stage assembly includes a rotating spindle configured to rotate the sample chamber relative to the optical axis of the lens assembly to allow selection of a rotating field of view. In some embodiments, the sample stage assembly allows for tilting and slight variation of the sample at the focal plane of the lens assembly and / or allows for both sample preparation and processing. In some embodiments, the sample stage assembly is configured to move the sample chamber away from the detection area and position different sample chambers in the detection area.

[0018] In some embodiments, the sample chamber is part of a centrifugal microfluidic biodisk, part of a capillary apparatus, or part of a flow cell apparatus. In some embodiments, the centrifugal microfluidic biodisk includes an elongated reservoir radially positioned at a first distance from the axis of rotation of the centrifugal microfluidic biodisk.

[0019] In another aspect, the Disclosure provides a system for analyzing a biological sample. The system includes an imaging device, such as any imaging device disclosed herein, configured to capture one or more images of the sample. The system also includes a control unit and a computing device. The control unit communicates with the imaging device by wire or wireless and is configured to control the imaging device to perform procedures for capturing data or images in a defined manner, for example, one or more light source assemblies, one or more detection assemblies, and / or other parts of the imaging device. The computing device communicates with the control unit by wire or wireless and processes one or more captured images of the sample.

[0020] In one embodiment, the control unit is a standalone unit. In another embodiment, the control unit is embedded in or integrated with the imaging device. In yet another embodiment, the control unit is embedded in or integrated with the computing device.

[0021] In further embodiments, the Disclosure provides a method for analyzing a biological sample. The method includes placing the sample at a working distance from the lens assembly of an imaging device, such as one of the imaging devices disclosed herein. The method also includes activating a first light source assembly of the imaging device to supply excitation light to the sample and detecting synchrotron radiation emitted from the sample by one or more detection assemblies of the imaging device. In some embodiments, the method also includes processing the detected synchrotron radiation to produce one or more sets of spectral and / or spatial responses, each set representing the spatial distribution of corresponding components within one or more components over an area of ​​the sample. The processing is performed on a computing device, such as a computing device disclosed herein.

[0022] The apparatus, systems, and methods of this disclosure have features and advantages that are evident from or described in more detail in the accompanying drawings incorporated herein and the embodiments for carrying out the inventions described herein, and together they help illustrate the specific principles of the exemplary embodiments of this disclosure.

[0023] The accompanying drawings are incorporated into and constitute part of this specification, illustrating one or more exemplary embodiments of the present disclosure and, together with modes for carrying out the invention, are useful in illustrating the principles and implementation of the exemplary embodiments of the invention. The accompanying drawings are not necessarily to scale. Certain design features of the invention as disclosed herein, such as certain dimensions, orientations, positions, and shapes, are partly determined by the specific intended use and environment. Furthermore, the illustrated components can be combined in any useful number and combinations. [Brief explanation of the drawing]

[0024] [Figure 1A] This is a schematic diagram showing a microscope imaging device according to some exemplary embodiments of the present disclosure. [Figure 1B]Schematic diagram showing a microscope imaging apparatus of FIG. 1A including an optical assembly according to some exemplary embodiments of the present disclosure. [Figure 1C] Schematic diagram showing a microscope imaging apparatus of FIG. 1A including an optical assembly according to an exemplary alternative embodiment of the present disclosure. [Figure 2] Schematic diagram showing a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 3] Schematic diagram showing a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 4] Schematic diagram showing a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 5] Schematic diagram showing a microscope system including a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 6] Flowchart showing a method for analyzing a biological sample according to some exemplary embodiments of the present disclosure. [Figure 7] Schematic diagram showing a lens assembly applicable to a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 8] Schematic diagram showing a microscope imaging apparatus according to some exemplary embodiments of the present disclosure. [Figure 9] Schematic diagram showing a centrifugal microfluidic biodisk according to some exemplary embodiments of the present disclosure.

Mode for Carrying Out the Invention

[0025] This disclosure provides portable microscope imaging devices, systems, and methods for analyzing biological samples such as bodily fluids (e.g., blood, plasma, urine). The microscope imaging devices of this disclosure are generally small, lightweight, and inexpensive. In certain embodiments, the microscope imaging devices of this disclosure are 1 / 10 to 1 / 20 the size of existing microscope imaging devices. The systems of this disclosure generally include a microscope imaging device that communicates with a computing device. Thus, the devices, systems, and methods provide the possibility of rapid screening, examination, and medical diagnosis not only in hospitals / clinics but also at home or other physical environments.

[0026] The reduction in size, weight, and cost of a microscope imaging device is achieved in part by the configuration of the microscope imaging device. For example, in certain embodiments, the microscope imaging device of the present disclosure includes a lens assembly having a short working distance, one or more light source assemblies, and one or more detection assemblies. One or more light source assemblies are configured to illuminate and / or excite a sample placed at the working distance of the lens assembly. One or more detection assemblies are configured to detect light from a sample, e.g., light emitted from the sample, light reflected by the sample, light scattered by the sample, or light transmitted through the sample. The lens assembly, at least one light source assembly, and at least one detection assembly are arranged to form an epitaxial configuration, i.e., both light from at least one light source assembly and light from the sample detected by at least one detection assembly pass through the same objective lens of the lens assembly.

[0027] The reduction in size, weight, and cost of the microscope imaging device is also partially achieved by using off-the-shelf components (e.g., mobile phones or consumer electronics). For example, in certain embodiments, the microscope imaging device of the Disclosure removes an off-the-shelf lens, reverses it, and uses the reversed off-the-shelf lens as the objective lens of the microscope imaging device. As a result, the short working distance of the objective lens of the microscope imaging device of the Disclosure is typically less than 5 millimeters (mm), less than 4 mm, less than 3 mm, less than 2 mm, less than 1.5 mm, less than 1.4 mm, less than 1.3 mm, less than 1.2 mm, less than 1.1 mm, or less than 1.0 mm. By using a reversed off-the-shelf lens as the objective lens, other off-the-shelf components can also be used, leading to further reductions in the overall size, weight, and cost of the microscope imaging device.

[0028] Referring here to the drawings, similar reference numerals refer to similar elements throughout, and Figures 1A to 1C show exemplary portable microscope imaging devices 100 according to several embodiments of the present disclosure. The imaging device 100 includes a lens assembly 120, a light source assembly 140, and a detection unit 180. The light source assembly 140 is configured to emit light when in operation. The lens assembly 120 includes a composite lens configured to optically communicate with the sample chamber 110 in the detection area (e.g., the working distance of the lens assembly, or the working distance of the objective lens 121 of the lens assembly). For example, the lens assembly 120 is configured to receive light emitted from the light source assembly 140 and guide the received light to the sample chamber 110 in the detection area. The lens assembly 120 is also configured to focus light from a sample placed in the sample chamber 110. The light focused from the sample includes light emitted from the sample, light scattered by the sample, light reflected by the sample, light transmitted through the sample, or any combination thereof. The lens assembly 120 is further configured to allow focused light to pass through for detection by the detection unit 180.

[0029] In some embodiments, the sample chamber is part of a centrifugal microfluidic biodisk, part of a capillary apparatus, or part of a flow cell apparatus. In some embodiments, the centrifugal microfluidic biodisk includes an elongated reservoir radially positioned at a first distance from the axis of rotation of the centrifugal microfluidic biodisk. As a non-limiting example, Figure 9 shows an exemplary centrifugal microfluidic biodisk 900. In some embodiments, the centrifugal microfluidic biodisk 900 includes an elongated reservoir 910 radially positioned at a first distance from the axis of rotation 920 of the centrifugal microfluidic biodisk.

[0030] In some embodiments, the imaging device 100 includes an optical assembly 130 that optically communicates with a lens assembly 120, a light source assembly 140, and a detection unit 180. The optical assembly 130 is configured to relay light emitted from the light source assembly 140 to the lens assembly 120, and then relay light from the sample, which has been focused and passed through the lens assembly 120, to the detection unit 180. For example, in some embodiments, the optical assembly 130 includes a dichroic mirror that reflects light at several wavelengths and transmits light at several other wavelengths. For example, the dichroic mirror has a passband that includes either (a) an excitation wavelength band emitted from a first light source, or (b) one or more emission wavelength bands. The dichroic mirror also has a stopband that includes either (a) an excitation wavelength band emitted from a first light source, or (b) the other of one or more emission wavelength bands.

[0031] As a non-limiting example, Figure 1B shows an optical assembly 130 including a dichroic mirror 131. The dichroic mirror 131 is positioned in the optical path between the lens assembly 120 and the light source assembly 140, and in the optical path between the lens assembly 120 and the detection assembly 180. In other words, the dichroic mirror 131 is in optical communication with the lens assembly, the light source assembly, and the detection assembly. The optical path between the lens assembly and the light source assembly is different from the optical path between the lens assembly and the detection assembly. The dichroic mirror 131 transmits light emitted from the first light source assembly (e.g., illumination light and / or excitation light) toward the lens assembly. The lens assembly directs the illumination light and / or excitation light to an area of ​​the sample placed at the working distance or focal plane of the lens assembly. The lens assembly focuses light from the sample (e.g., light emitted from the sample, light scattered by the sample, light reflected by the sample, and / or light transmitted through the sample) and passes the focused light through the dichroic mirror 131. The dichroic mirror 131 reflects the light that has been focused and passed through by the lens assembly back to the detection assembly.

[0032] As another non-limiting example, Figure 1C shows an optical assembly 130 including a dichroic mirror 132. Similar to the dichroic mirror 131, the dichroic mirror 132 is positioned in the optical path between the lens assembly and the light source assembly, and in the optical path between the lens assembly and the detection assembly, which is different from the optical path between the lens assembly and the light source assembly. Unlike the dichroic mirror 131, the dichroic mirror 132 reflects light emitted from the first light source assembly to the lens assembly and transmits the light that has been focused and passed through the lens assembly toward the detection assembly.

[0033] In various embodiments, the lens assembly 120 has (i) an optical path of less than approximately 15 mm, (ii) a working distance of approximately 5 mm or less, (iii) a weight of approximately 50 grams or less, and (iv) more than three optical elements, or any combination thereof. For example, as a non-limiting example, Figure 7 shows a lens assembly 120 including more than three optical elements. Of the more than three optical elements, the first optical element 710 is closest to the object-side plane of the lens assembly, and the second optical element 720 is furthest from the object-side plane of the lens assembly. The first optical element 710 has a first effective aperture diameter (generally denoted as D1). The second optical element 720 has a second effective aperture diameter (generally denoted as D2). The first effective aperture diameter D1 is greater than or equal to the second effective aperture diameter D2. For example, in some embodiments, the D1 / D2 ratio is about 1, about 1.1, about 1.2, about 1.3, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.25, about 2.5, about 2.75, or about 3. In some embodiments, the D1 / D2 ratio is at least 1, at least 1.1, at least 1.2, at least 1.3, at least 1.4, at least 1.5, at least 2, at least 2.5, at least 3, at least 3.5, at least 4, at least 4.5, or at least 5.

[0034] In some embodiments, the ratio of D1 / D2 is 1-10, 1-5, 1-4, 1-3, 1-2.5, 1-2.25, 1-2, 1-1.75, 1-1.5, 1-1.25, 1-1.1, 1.1-10, 1.1-5, 1.1-4, 1.1-3, 1.1-2.5, 1.1-2.25, 1.1-2, 1.1-1.75, 1.1-1.5, 1.1-1.25, 1.25-10, 1.25-5, 1.25-4, 1.25-3, 1.25-2.5, 1.25-2.25, 1.25-2, 1.25-1.75, 1.25-1.5, 1.5-10, 1.5- 5, 1.5~4, 1.5~3, 1.5~2.5, 1.5~2.25, 1.5~2, 1.5~1.75, 1.75~10, 1.75~5, 1.75~4, 1.75~3, 1.75~2.5, 1.75~2.25, 1.75~2, 2~10, 2~5, 2~4, 2~3, 2~2.5, 2~2.25, 2.5~10, 2.5~7.5, 2.5~5, 2.5~5, 2.5~3, 3~10, 3~7.5, 3~5, 3~4, 4~10, 4~7.5, 4~5, 5~10, 5~7.5, 7.5~5, or any other range starting with 1 or more and ending with 10 or less.

[0035] In some embodiments, the lens elements in front of the object plane (e.g., adjacent) deviate significantly from a spherical shape. The refractive power of the lens at the center of the lens is also shown, whether positive or negative. However, the first two lens elements have a strong field-of-view dependent effect due to their (locally) very different refractive powers and beam deflection. Due to local curvature variations, the refractive power changes along each field-of-view dependent optical path. Positive (+), neutral (o), and negative (-) forces are illustrated for fields of view near lens elements 1, 2, and 3. This enables better aberration correction of the objective lens and a wider field of view in a smaller form factor. For example, in some embodiments, the lens assembly 120 has a total length (generally represented as L) and (generally,

number

[0036]

number

number

[0037] The compactness factor of a lens assembly may also be calculated by multiplying the r value by the magnification of the lens assembly. As a non-limiting example, Table I below lists the compactness factors of several exemplary lens assemblies with a total length L of approximately 10 mm, and the diagonal dimensions

number

number

[0038] Therefore, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 4x, and a compactness factor of 10 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 4x, and a compactness factor of 8 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 4x, and a compactness factor of 6 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 4x, and a compactness factor of 5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 4x, and a compactness factor of 4 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0039] Similarly, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 8x, and a compactness factor of 17.5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 8x, and a compactness factor of 15 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 8x, and a compactness factor of 12.5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 8x, and a compactness factor of 10 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 8x, and a compactness factor of 8 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0040] Similarly, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 10x, and a compactness factor of 25 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 10x, and a compactness factor of 20 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 10x, and a compactness factor of 15 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 10x, and a compactness factor of 12.5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 10x, and a compactness factor of 10 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0041] Therefore, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 20 times, and a compactness factor of 40 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 20 times, and a compactness factor of 30 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 20 times, and a compactness factor of 25 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 20 times, and a compactness factor of 20 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 20 times, and a compactness factor of 17.5 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0042] Therefore, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 40 times, and a compactness factor of 80 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 40 times, and a compactness factor of 60 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 40 times, and a compactness factor of 50 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 40 times, and a compactness factor of 40 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 8 mm to 12 mm, a magnification of at least 40 times, and a compactness factor of 35 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0043] Similarly, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 4x, and a compactness factor of 8 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 4x, and a compactness factor of 6 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 4x, and a compactness factor of 5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 4x, and a compactness factor of 4 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 4x, and a compactness factor of 2 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0044] Similarly, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 8x, and a compactness factor of 12 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 8x, and a compactness factor of 10 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 8x, and a compactness factor of 8 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 8x, and a compactness factor of 6 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 8x, and a compactness factor of 5 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0045] Similarly, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 10x, and a compactness factor of 14 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 10x, and a compactness factor of 12 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 10x, and a compactness factor of 10 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 10x, and a compactness factor of 8 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 10x, and a compactness factor of 6 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0046] Therefore, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 20 times, and a compactness factor of 25 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 20 times, and a compactness factor of 20 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 20 times, and a compactness factor of 17.5 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 20 times, and a compactness factor of 15 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 20 times, and a compactness factor of 12.5 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0047] Therefore, in some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 40 times, and a compactness factor of 50 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 40 times, and a compactness factor of 40 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 40 times, and a compactness factor of 35 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 40 times, and a compactness factor of 30 or less. In some embodiments, the lens assembly used in the imaging device described herein has (i) an overall length of 6 mm to 8 mm, a magnification of at least 40 times, and a compactness factor of 25 or less. In some such embodiments, the lens assembly has a field of view of 15 mm or less. In some such embodiments, the lens assembly has a field of view of 12 mm or less. In some such embodiments, the lens assembly has a field of view of 10 mm or less. In some such embodiments, the lens assembly has a field of view of 9 mm or less. In some such embodiments, the lens assembly has a field of view of 8 mm or less. In some such embodiments, the lens assembly has a field of view of 7 mm or less. In some such embodiments, the lens assembly has a field of view of 6 mm or less. In some such embodiments, the lens assembly has a field of view of 5 mm or less.

[0048] However, the present invention is not limited to these. The imaging apparatus of the present invention may include other lens assemblies. For example, in some embodiments, the lens assembly of the present invention is identical or similar to that disclosed by Blahnik, et al., “Smartphone imaging technology and its applications”, Advanced Optical Technologies, vol.10, no.3, 2021, pp.145-232, and the whole thereof is incorporated herein, regardless of its purpose.

[0049] In some embodiments, the lens assembly includes a composite lens having an optical axis and an image plane through which the optical axis passes. The composite lens includes a plastic lens barrel surrounding the optical axis. The plastic lens barrel includes an image-side portion through which the optical axis passes and an object-side aperture. When in use, the sample is placed on the limited focal plane of the composite lens at a distance expressed in the design as the rear focal length. In some embodiments, the composite lens includes two imaging lenses having two different rear focal lengths. The object side uses the imaging lens with the shorter rear focal length, and the image side uses the imaging lens with the longer rear focal length. The effective magnification of the entire imaging device is determined by the ratio of the rear focal lengths of the two imaging lenses. This configuration allows for the use of various imaging lens designs that can result in magnifications of 1x, 2x, 5x, 10x, 15x, 20x, or greater. In some embodiments, the composite lens includes 2 to 10 lenses.

[0050] In certain embodiments, the lens assembly 120 has a short working distance. For example, in some embodiments, a general-purpose lens (e.g., from a mobile phone or consumer electronics) is taken out, reversed, and used as the objective lens 121 of the lens assembly. A typical general-purpose lens package has more than three optical elements and a short rear focal length (e.g., less than 2 mm) to focus an image onto the image sensor. It is usually less than 15 mm thick, less than 15 mm in diameter, and weighs less than 50 grams (g). General-purpose lenses are also inexpensive, costing only a few dollars in the current market. Therefore, by using a reversed general-purpose lens as the objective lens, the microscope imaging apparatus of this disclosure can achieve a short working distance substantially the same as the rear focal length (e.g., less than 2 mm) of a general-purpose lens, while reducing overall size, weight, and cost.

[0051] In some embodiments, the working distance of the lens assembly 120 is less than 5 mm, less than 4 mm, less than 3 mm, or less than 2 mm. In some embodiments, the working distance is less than approximately 1.5 mm, less than 1.4 mm, less than 1.3 mm, less than 1.2 mm, less than 1.1 mm, or less than 1.0 mm. In some embodiments, the working distance is approximately 1.0 mm, approximately 1.2 mm, approximately 1.4 mm, or approximately 1.6 mm.

[0052] In some embodiments, the objective lens 121 or lens assembly 120 is less than 15 mm thick and less than 15 mm in diameter. In some embodiments, the thickness of the objective lens 121 or lens assembly 120 is less than 14 mm, less than 13 mm, less than 12 mm, less than 11 mm, less than 10 mm, less than 9 mm, less than 8 mm, less than 7 mm, less than 6 mm, less than 5 mm, less than 4 mm, or less. In one embodiment, the objective lens 121 or lens assembly 120 is about 5 mm thick and about 7 mm in diameter.

[0053] In some embodiments, the weight of the objective lens 121 or lens assembly 120 is less than 50g, less than 45g, less than 40g, less than 35g, less than 30g, less than 25g, less than 20g, less than 15g, less than 10g, less than 9g, less than 8g, less than 7g, less than 6g, or less than 5g. In one embodiment, the weight of the objective lens 121 or lens assembly 120 is about 10g. In another embodiment, the weight of the objective lens 121 or lens assembly 120 is about 8g. In yet another embodiment, the weight of the objective lens 121 or lens assembly 120 is about 6g.

[0054] Furthermore, reversing a standard lens and using the reversed standard lens as the objective lens allows for the use of other standard components, leading to further reductions in the overall size, weight, and cost of the microscope imaging device. For example, in some embodiments, the imaging device includes an actuator 160, such as a voice coil actuator (VCA) or voice coil motor (VCM), coupled to or integrated with the lens assembly, to adjust the focus of the objective lens or lens assembly. In some embodiments, the actuator 160 is a standard autofocus VCA or VCM.

[0055] In some embodiments, the size of the actuator 160 (e.g., vca or vcm) is less than 30 mm in diameter and less than 30 mm in thickness. In some embodiments, the diameter of the actuator is less than 32 mm, less than 30 mm, less than 28 mm, less than 26 mm, less than 24 mm, less than 22 mm, less than 20 mm, or less. In some embodiments, the actuator can move or fine-tune the objective lens or lens assembly within a range of about ±1.2 mm, about ±1.1 mm, about ±1.0 mm, about ±0.9 mm, or about ±0.8 mm.

[0056] In some embodiments, the sample to be detected is mounted on a sample stage assembly 170 configured to position the sample chamber in the detection area. For example, in some embodiments, the sample stage assembly is configured to move the sample chamber relative to the lens assembly. In some embodiments, the sample stage assembly is configured to allow selection of a rotating field of view (FOV). For example, in some embodiments, the sample stage assembly includes a rotating spindle configured to rotate the sample chamber relative to the optical axis of the lens assembly to allow selection of a rotating field of view. In one embodiment, the sample stage assembly includes a single rotating spindle for adjusting the FOV, for example, by allowing a consistent tilt and slight variation of the sample at the focal plane of the lens assembly. By using a single rotating spindle, the sample stage assembly of the present disclosure has fewer moving parts than a conventional sample stage that moves along two axes. As a result, the imaging device of the present disclosure is simpler and less expensive. Furthermore, the sample stage assembly of the present disclosure allows for sample preparation and sample handling in the same structure as imaging without requiring additional pumps, valves, or manual intervention. In some embodiments, the sample stage assembly is configured to move the sample chamber away from the detection area and position different sample chambers in the detection area.

[0057] The light source assembly 140 is configured to emit light for irradiating a sample and / or for exciting one or more components of the sample. For example, in some embodiments, the light source assembly 140 is configured to emit irradiation light, for example, irradiation light in which at least a portion of the light is suitable for irradiating a sample but cannot excite any component in the sample. In some embodiments, the light source assembly 140 is configured to emit excitation light, for example, excitation light in which at least a portion of the light can excite one or more components of the sample. In some embodiments, the light source assembly 140 is configured to emit light including both irradiation light and excitation light.

[0058] In some embodiments, the light source assembly 140 is configured to emit light at one or more wavelengths or in one or more wavelength ranges. The wavelength range may be broad or narrow (e.g., broadband or narrowband, and the term “wavelength range” is used synonymously with the term “wavelength band”). Different wavelength ranges may be continuous, discontinuous, or overlapping. For example, in a non-limiting example, the light source assembly 140 may be configured to emit light in a broad, continuous wavelength range, for example, visible light in the wavelength range of 400 nanometers (nm) to 700 nm. In another non-limiting example, the light source assembly 140 may be configured to emit light containing excitation light in one or more excitation wavelength ranges to excite one or more components of a sample. In a further non-limiting example, the light source assembly 140 may include a light source that emits light over a relatively broad range and one or more filters (e.g., bandpass filters) that filter the emitted light to produce excitation light in one or more excitation wavelength ranges to excite one or more components of a sample. In some embodiments, the first light source assembly is configured to emit light in a single narrow wavelength band. In some embodiments, the full width at half maximum (FWHM) of the single narrow wavelength band is 50 nm or less, 25 nm or less, 10 nm or less, or 5 nm or less.

[0059] The light source assembly 140 includes a light source 141. The light source 141 may include one or more laser diodes, one or more light-emitting diodes (LEDs), or any combination thereof. LEDs are preferred over laser diodes because laser diodes can cause excessive heating and photobleaching. Furthermore, laser diodes on the current market may not produce light at specific desired wavelengths or wavelength ranges for analyzing particular biological samples. In comparison, LEDs typically generate less heat and can provide more uniform light and consistent light output over time. In some embodiments, a single LED can supply sufficient light power to the sample plane with less heating. Thus, the chance of sample damage and burnout is reduced.

[0060] In one embodiment, the light source 141 consists of a single LED. In some embodiments, the light source 141 includes multiple LEDs, for example, two, three, four, five, six, seven, eight, nine, ten, or more than ten LEDs. In one embodiment, the multiple LEDs are arranged as a single point light source. In another embodiment, the multiple LEDs are arranged as a two-dimensional light source, for example, in an array on a common plane or forming a circle. In yet another embodiment, the multiple LEDs are arranged as a three-dimensional light source, for example, not on a common plane. In one embodiment, all of the multiple LEDs emit light in the same wavelength band. In another embodiment, the multiple LEDs include at least one LED configured to emit light in a first narrow wavelength band and at least one LED configured to emit light in a second narrow wavelength band different from the first narrow wavelength band. In further embodiments, each of the multiple LEDs emits light in a different wavelength band.

[0061] For example, in some embodiments, the microscope imaging apparatus of the present disclosure is configured to analyze biological samples such as bodily fluids (e.g., blood samples, plasma samples, buffy coat samples, urine samples). Blood samples generally include blood cells (e.g., red blood cells, white blood cells, and platelets), and urine samples generally include analytes (e.g., various types of crystals present in urine). In such embodiments, the light source 141 is configured to emit light in one or more wavelength ranges corresponding to cell staining and / or dyes (e.g., nucleic acids, organelles, or staining of other cellular structures or substructures and / or labels bound to cell surface markers). In some embodiments, the light source assembly 141 is configured to emit excitation light in one or more excitation wavelength ranges to excite one or more components of the sample (e.g., dyes, fluorescent labels). When excited, one or more components of the sample emit synchrotron radiation in one or more emission wavelength ranges.

[0062] In some embodiments, the light source 141 is configured to emit light containing excitation light in one or more narrow wavelength bands. One or more narrow wavelength bands may be selected from the group consisting of 405 nanometers (nm), 460 nm, 470 nm, 520 nm, and 638 nm. In one embodiment, light from a first narrow wavelength band excites a first component in the sample (e.g., DNA or a dye that stains DNA), and light from a second narrow wavelength band excites a second component in the sample (e.g., RNA or a dye that stains RNA). Once excited, the first component emits light (e.g., green light) in a first emission wavelength range, and the second component emits light (e.g., red light) in a second emission wavelength range. In an alternative embodiment, the light source 141 is configured to emit light containing excitation light in a single narrow wavelength band that excites both the first and second components (e.g., the same dye that stains both DNA and RNA). When excited, the first component emits light (e.g., green light) in a first emission wavelength range, and the second component emits light (e.g., red light) in a second emission wavelength range.

[0063] In some embodiments, the light source assembly 140 includes additional optical components. For example, in some embodiments, the light source assembly 140 includes a collimating lens 142, a diffuser 143, a baffle 144, or any combination thereof. Additional or optional components are included to collimate the light emitted from the light source 141, soften the light, and / or prevent the light from scattering in unwanted directions.

[0064] The detection unit 180 includes one or more detection assemblies. For example, Figure 1A shows that the detection unit 180 includes detection assembly 181 and detection assembly 184. Detection assembly 181 and detection assembly 184 can be configured to be identical, similar, or different from one another. In some embodiments, detection assembly 181 includes a sensor 182 and a tube lens 183 that receives light and focuses the light onto the sensor 182. In some embodiments, the effective focal length of the tube lens is less than 50 mm, less than 45 mm, less than 40 mm, less than 35 mm, or less than 30 mm. The use of a tube lens enables more advanced optical manipulation techniques. Similarly, in some embodiments, detection assembly 184 includes a sensor 185 and a tube lens 186 that receives light and focuses the light onto the sensor 185. In one embodiment, the sensor 185 and tube lens 185 of detection assembly 184 are substantially the same as the sensor 182 and tube lens 183 of detection assembly 181.

[0065] In some embodiments, the sensor 181 or 185 is a general-purpose sensor (e.g., a consumer electronics camera sensor) and is compatible with the Mobile Industry Processor Camera Serial Interface standard for high-speed communication between the sensor and the host processor. In some embodiments, the sensor is a charge-coupled device (CCD), etc. In some embodiments, the cross-sectional or diagonal dimension of the sensor is less than 20 mm, less than 19 mm, less than 18 mm, less than 17 mm, less than 16 mm, or less than 15 mm. In some embodiments, the sensor includes at least 500 pixels. In some embodiments, the sensor includes at least 1,000 pixels, at least 2,500 pixels, at least 5,000 pixels, at least 10,000 pixels, at least 25,000 pixels, at least 50,000 pixels, at least 100,000 pixels, at least 250,000 pixels, at least 500,000 pixels, at least 1,000,000 pixels, at least 2,500,000 pixels, at least 5,000,000 pixels, or more pixels. Thus, the sensor can detect the spatial response of a sample in response to irradiation and / or excitation.

[0066] In some embodiments, detection assemblies 181 and 184 are configured or arranged to detect light in different wavelength ranges. For example, in some embodiments, the imaging device or detection unit includes a dichroic mirror 187 positioned in the optical path to receive light from an optical assembly 130. The dichroic mirror 187 has a passband including a first portion of one or more emission wavelength bands and a stopband including a second portion of one or more emission wavelength bands. In some embodiments, the dichroic mirror 187 transmits light in one or more wavelength ranges and reflects light in one or more different wavelength ranges. In non-limiting examples, the dichroic mirror 187 transmits green light and reflects red light, or vice versa. One of detection assemblies 181 and 184 is configured or arranged to detect light transmitted through the dichroic mirror 187, and the other of detection assemblies 181 and 184 is configured or arranged to detect light reflected by the dichroic mirror 187. As an example, Figure 1A shows that detection assembly 181 detects light transmitted through the dichroic mirror 187, and detection assembly 184 detects light reflected by the dichroic mirror 187. Naturally, detection assembly 181 can be configured or positioned to detect light reflected by the dichroic mirror 187, and detection assembly 184 can be configured or positioned to detect light transmitted through the dichroic mirror 187.

[0067] By using dichroic mirrors and multiple detection assemblies, the need for a filter wheel for channel separation (e.g., separation of light at different wavelengths or wavelength bands) is eliminated. Furthermore, faster image capture is possible without the delay caused by moving the filter wheel, which inherently adds time between image captures. In addition, it provides the possibility of sequential or simultaneous measurement of multiple channels as needed. Moreover, additional dichroic mirrors(s), bandpass filters, and / or detection assemblies can be used for further channel separation.

[0068] In some embodiments, the imaging device includes other additional, optional, or alternative components. For example, in some embodiments, the imaging device or detection unit includes a filter 188 positioned in the optical path between the optical assembly 130 and the dichroic mirror 187 to remove noise or unwanted light, etc. In one embodiment, the filter 188 includes a long-pass filter that reflects short-wavelength light while transmitting or passing long-wavelength light through.

[0069] Referring to Figure 8, in some embodiments, alternatively, optionally, or additionally, the imaging device includes at least one splitter and a plurality of filters. One or more detection assemblies include a plurality of detection assemblies. At least one splitter is located in a second optical path between a first dichroic mirror and the plurality of detection assemblies and is configured to divide one or more emission wavelength bands into a plurality of parts. Each of the plurality of filters is located between at least one splitter and a corresponding detection assembly in the plurality of detection assemblies and has its own passband in a plurality of passbands, filtering the corresponding part in the plurality of parts of one or more emission wavelength bands. For example, as a non-limiting example, Figure 8 shows an imaging device including a splitter 810 and a plurality of filters 820-1, 820-2, etc. The splitter 810 is located in a second optical path between a dichroic mirror 131 of optical assembly 130 and a plurality of detection assemblies 830-1, 830-2, etc. The splitter 810 is configured to divide one or more emission wavelength bands (e.g., light received from the optical assembly 130) into n parts, where n is an integer greater than 1. The filter 820-1 is positioned between the splitter 810 and the detection assembly 830-1 and has a first passband for filtering a first part of one or more emission wavelength bands. The detection assembly 830-1 includes a two-dimensional photodetector (e.g., sensor 181 or 185) configured to acquire an image of the first part of one or more emission wavelength bands after passing through the filter 820-1. Similarly, the filter 820-2 is positioned between the splitter 810 and the detection assembly 830-2 and has a second passband for filtering a second part of one or more emission wavelength bands. The detection assembly 830-2 includes a two-dimensional photodetector (e.g., sensor 181 or 185) configured to acquire an image of the second part of one or more emission wavelength bands after passing through the filter 820-2.

[0070] In some embodiments, the imaging device includes an additional or optional light source assembly 150. The light source assembly 150 may be configured identically to, similarly to, or differently from the light source assembly 140. The light source assembly 150 may be configured to emit illumination and / or excitation light. It may also be positioned to supply illumination and / or excitation light to the sample from the same side or a different side relative to the lens assembly. For example, in some embodiments, the sample is positioned so that a first side (e.g., the bottom side of the sample in the figure) faces the lens assembly. The light source assembly 150 may be positioned to supply illumination and / or excitation light reaching the first side of the sample vertically or at an angle. The light source assembly 150 may also be positioned to supply illumination and / or excitation light reaching a second side of the sample vertically or at an angle. As an example, the light source assembly 150 is shown to supply illumination and / or excitation light from a different side relative to the lens assembly.

[0071] The use of light source assemblies 140 and 150 enables epifluorescence imaging techniques in combination with other imaging techniques such as side scattering, dark-field, and / or bright-field imaging techniques. For example, in some embodiments, light source assembly 140 is configured to supply light containing excitation light to excite one or more components of a sample that, when excited, emit fluorescence at one or more emission wavelengths or emission wavelength ranges. Light source assembly 150 includes one or more of the bright-field light source 152, the dark-field light source 153, and / or side-scatter light source 151. Light source assembly 150 emits light containing illumination light that propagates to different sides of the sample relative to the lens assembly. The sample reflects, scatters, absorbs, and / or transmits the illumination light from light source assembly 150. The lens assembly focuses at least a portion of the light emitted from the sample, scattered by the sample, and / or transmitted through the sample, and passes the focused light for detection by the detection unit 180.

[0072] Brightfield light sources are generally positioned approximately 45 degrees relative to the irradiation area of ​​the sample or the optical axis of the lens assembly. ° ~about 90 ° The light source is positioned to illuminate the sample at an angle of 0. ° ~about 45 ° The light source assembly is positioned to irradiate the sample at an angle. In some embodiments, the light source assembly 150 includes other optional or additional components such as a diffuser 154 and one or more apertures 155. In some embodiments, the diffuser 154 is configured to soften or diffuse light from a bright-field, dark-field, or side-scatter light source. In some embodiments, one or more apertures 155 are configured to allow light from a bright-field, dark-field, or side-scatter light source to pass through.

[0073] In some embodiments, the bright-field light source 152 includes one or more LEDs, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more than ten LEDs. In some embodiments, the dark-field light source 153 includes one or more LEDs, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more than ten LEDs. In some embodiments, the side-scatter light source 151 includes one or more LEDs, for example, one, two, three, four, five, six, seven, eight, nine, ten, or more than ten LEDs. In one embodiment, each of the bright-field light source, dark-field light source, and scattering light source includes one or more LEDs.

[0074] While Figure 1A shows a microscope imaging device including two light source assemblies and two detection assemblies, it should be noted that the microscope imaging device of the present disclosure may include one, two, three, four, or more light source assemblies for illuminating and / or exciting a sample, and one, two, three, four, or more detection assemblies for detecting light from the sample (e.g., light emitted from the sample, light reflected by the sample, light scattered by the sample, light transmitted through the sample, or any combination thereof). In various exemplary embodiments, the lens assembly, at least one light source assembly, and at least one detection assembly are arranged to form an epitaxial configuration, i.e., both light from at least one light source assembly and light from the sample detected by at least one detection assembly pass through the same objective lens of the lens assembly.

[0075] For example, in a non-limiting example, Figure 2 shows an exemplary microscope imaging apparatus 200 comprising a single light source assembly (e.g., light source assembly 140) and a single detection assembly (e.g., detection assembly 181 or 184) according to some exemplary embodiments of the present disclosure. In the illustrated embodiment, an optical assembly 130 relays light emitted from the light source assembly to a lens assembly 120, and then relays light from the sample, which has been focused and passed through the lens assembly 120, to a single detection assembly.

[0076] As another non-limiting example, Figure 3 shows an exemplary microscope imaging apparatus 300 comprising two light source assemblies (e.g., light source assembly 140 and an additional or optional light source assembly 150) and a single detection assembly (e.g., detection assembly 181 or 184), according to some exemplary embodiments of the present disclosure. The imaging apparatus 300 is similar to the imaging apparatus 200 except for the presence of the additional or optional light source assemblies. In some embodiments, the light source assembly 140 supplies excitation light to a sample, and the sample generates emission light in response to the excitation light. The light source assembly 150 generates illumination light to the sample, and the sample reflects, scatters, absorbs, and / or transmits the light from the light source assembly 150. A lens assembly focuses at least a portion of the light emitted from the sample, scattered by the sample, and / or transmitted through the sample, and passes the focused light through an optical assembly 130. The optical assembly 130 relays the light from the sample, which has been focused and passed through by the lens assembly 120, to a single detection assembly.

[0077] As a further non-limiting example, Figure 4 shows an exemplary microscope imaging device 400 comprising a single light source assembly (e.g., light source assembly 140) and two detection assemblies (e.g., detection assembly 181 and detection assembly 184), according to some exemplary embodiments of the present disclosure. The imaging device 400 is similar to the imaging device 100 except for the absence of additional or optional light source assemblies. In some embodiments, the light source assembly 140 is configured to supply excitation light to a sample, which emits light in response to the excitation light. A lens assembly focuses at least a portion of the light emitted from the sample and passes the focused light through an optical assembly 130 for detection by the detection assembly. In some embodiments, the light source assembly 140 is configured to supply illumination light to a sample, which reflects, scatters, absorbs, and / or transmits light from the light source assembly 140. A lens assembly focuses at least a portion of the light scattered by and / or reflected by the sample and passes the focused light through an optical assembly 130 for detection by the detection assembly. In some embodiments, the light source assembly 140 is configured to supply both excitation light and irradiation light to the sample, and the lens assembly collects at least a portion of the light emitted from the sample, scattered by the sample, and / or reflected by the sample, and passes the collected light to the optical assembly 130 for detection by the detection assembly.

[0078] The portable microscope imaging devices of this disclosure (e.g., imaging devices 100, 200, 300, or 400) are generally small, having dimensions of less than 25 centimeters (cm) in length, less than 25 cm in width, and less than 25 cm in height. In some embodiments, the portable microscope imaging devices of this disclosure (e.g., imaging devices 100, 200, 300, or 400) are about 10 times, about 12 times, about 14 times, about 16 times, about 18 times, about 20 times, about 22 times, about 24 times, about 26 times, about 28 times, or about 30 times smaller than some existing microscope imaging devices. In some embodiments, the portable microscope imaging devices of this disclosure are more than 30 times smaller than existing microscope imaging devices. For example, in some embodiments, the overall size of the portable microscope imaging device of this disclosure is less than about 200 mm in length, about 25 mm in width, and about 35 mm in height. In some embodiments, the overall dimensions of the portable microscope imaging device of this disclosure are less than approximately 200 mm in length, 22 mm in width, and 33 mm in height. In some embodiments, the overall dimensions of the portable microscope imaging device of this disclosure are less than approximately 180 mm in length, 20 mm in width, and 30 mm in height. In some embodiments, the overall length of the portable microscope imaging device of this disclosure is less than 200 mm, less than 190 mm, less than 180 mm, less than 170 mm, less than 160 mm, less than 150 mm, less than 140 mm, less than 130 mm, less than 120 mm, or less. In some embodiments, the overall width of the portable microscope imaging device of this disclosure is less than 22 mm, less than 21 mm, less than 20 mm, less than 19 mm, less than 18 mm, less than 17 mm, less than 16 mm, less than 15 mm, or less. In some embodiments, the total height of the portable microscope imaging device of the Disclosure is less than 35 mm, less than 34 mm, less than 33 mm, less than 32 mm, less than 31 mm, less than 30 mm, less than 29 mm, less than 28 mm, less than 27 mm, less than 26 mm, less than 25 mm, or less. In some embodiments, the weight of the portable microscope imaging device of the Disclosure is less than 300 mg (g), less than 280 g, less than 260 g, less than 240 g, less than 220 g, less than 200 g, less than 180 g, less than 160 g, less than 150 g, or less.

[0079] In some embodiments, the F-number of the portable microscope imaging device of the Disclosure is approximately f / 2.0, approximately f / 2.1, approximately f / 2.2, approximately f / 2.3, approximately f / 2.4, or approximately f / 2.5. In some embodiments, the F-number of the portable microscope imaging device of the Disclosure is greater than approximately f / 2.0, greater than approximately f / 2.5, greater than approximately f / 3.0, greater than approximately f / 3.5, greater than f / 4.0, greater than approximately f / 4.5, greater than approximately f / 5.0, or higher. In some embodiments, the magnification of the portable microscope imaging device of the Disclosure is less than approximately 10x. In some embodiments, the magnification of the portable microscope imaging device of the Disclosure is less than approximately 5x, less than approximately 10x, less than approximately 15x, less than approximately 20x, less than approximately 25x, less than approximately 30x, less than approximately 40x, less than approximately 50x, less than approximately 75x, less than approximately 100x, or higher.

[0080] Referring now to Figure 5, a block diagram is shown illustrating a system 500 for analyzing a biological sample according to several embodiments of the present disclosure. The system 500 includes an imaging device 560, a control unit 550, and a computing device 501. The imaging device 560 may be any of the imaging devices 100, 200, 300, and 400 disclosed herein.

[0081] The control unit can be a standalone unit. The control unit can also be embedded in or integrated with the imaging device 560 or the computing device 501. In some embodiments, the control unit 550 is configured to control the imaging device 560 so that detection (e.g., capturing data or images) can be performed in a desired or defined manner. For example, in an embodiment in which the imaging device includes a light source assembly 140, the control unit is configured to selectively activate the light source assembly 140 or any individual light sources within the light source assembly 140 (e.g., one or more LEDs). In an embodiment in which the imaging device includes a light source assembly 150, the control unit is configured to selectively activate the light source assembly 150 or any individual light sources within the light source assembly 150 (e.g., one or more LEDs of a bright-field / dark-field / side-scatter light source or a bright-field / dark-field / side-scatter light source). In embodiments where the imaging device includes a light source assembly 140 and a light source assembly 150, the control unit is configured to selectively operate the light source assembly, the second light source assembly, any individual light source in the first light source assembly, any individual light source in the second light source assembly, or any combination thereof, in a defined sequence. In some embodiments, the control unit controls an autofocus VCA or VCM, one or more light source assemblies, and one or more detection assemblies to perform the entire data or image capture procedure in a defined manner (e.g., sequentially, simultaneously, or sequentially in a defined sequence).

[0082] In some embodiments, the computing device 501 is configured identically or similarly to the computing device disclosed in U.S. Patent Application No. 17 / 371,746, the contents of which are incorporated herein by reference in their entirety. For example, in some embodiments, the computing device 501 includes a network interface 504. In some embodiments, the network interface 504 interconnects components within the computing device 501, as well as optional external systems and devices, via one or more communication networks (e.g., via an optional network communication module 518). In some embodiments, the network interface 504 optionally provides communication via the Internet, one or more local area networks (LANs), one or more wide area networks (WANs), other types of networks, or a combination of such networks.

[0083] Examples of networks include the World Wide Web (WWW), intranets and / or wireless networks, such as cellular networks, wireless local area networks (LANs) and / or large area networks (MANs), and other devices using wireless communication. Wireless communication can be optionally selected from Global System for Mobile Communications (GSM), Enhanced Data GSM Environment (EDGE), High-Speed ​​Downlink Packet Access (HSDPA), High-Speed ​​Uplink Packet Access (HSUPA), Evolution, Data Only (EV-DO), HSPA, HSPA+, Dual Cell HSPA (DC-HSPADA), Long-Term Evolution (LTE), Near Field Communication (NFC), Wideband Code Division Multiple Access (W-CDMA), Code Division Multiple Access (CDMA), Time Division Multiple Access (TDMA), Bluetooth, Wireless Fidelity (Wi-Fi) (e.g., IEEE 802.11a, IEEE 802.11ac, IEEE 802.11ax, IEEE 802.11b, IEEE 802.11g and / or IE Using any of several communication standards, protocols, and technologies, including EE802.11n), Voice over Internet Protocol (VoIP), Wi-MAX, email protocols (e.g., Internet Message Access Protocol (IMAP) and / or Post Office Protocol (POP)), instant messaging (e.g., Extensible Messaging and Presence Protocol (XMPP), Session Initiation Protocol for Instant Messaging and Presence Leveling Extensions (SIMPLE), Instant Messaging and Presence Service (IMPS)), and / or Short Message Service (SMS), or any other suitable communication protocol, including communication protocols not yet developed as of the filing date of this specification.

[0084] In some embodiments, the computing device 501 includes one or more processing units (CPUs) 502 (e.g., a processor, processing core, etc.), one or more network interfaces 504, a user interface 506 (optionally) including a user display 508 and an input system 510 (e.g., an input / output interface, keyboard, mouse, etc.), memory (e.g., non-persistent memory 511, persistent memory 512), and one or more communication buses 514 for interconnecting the above components. The one or more communication buses 514 optionally include circuits (sometimes called chipsets) for interconnecting and controlling communication between system components. Non-persistent memory 511 typically includes high-speed random-access memory such as DRAM, SRAM, DDR RAM, ROM, EEPROM, and flash memory, while persistent memory 512 typically includes CD-ROMs, digital multipurpose discs (DVDs) or other optical storage, magnetic cassettes, magnetic tapes, magnetic disk storage or other magnetic storage devices, magnetic disk storage devices, optical disk storage devices, flash memory devices, or other non-volatile solid-state storage devices. The persistent memory 512 optionally includes one or more storage devices located remotely from the CPU(s) 502. The non-volatile memory devices(s) in the persistent memory 512 and non-persistent memory 512 include non-temporary computer-readable storage media. In some embodiments, the non-persistent memory 511 or alternative non-temporary computer-readable storage media sometimes work in conjunction with the persistent memory 512 to store the following programs, modules, and data structures, or subsets thereof: • Operating systems 516 (e.g., embedded operating systems such as ANDROID, iOS, DARWIN, RTXC, LINUX, UNIX, OS X, WINDOWS, or VxWorks) that include procedures for handling various basic system services and performing hardware-dependent tasks. • An optional network communication module (or instruction) 518 for connecting computing device 501 to other devices and / or communication networks. • Image data store 520 for storing pixelated images, such as digital images, for one or more samples. • Image processing module 530 including one or more algorithms or models for analyzing data or images. • An optional report module 540 for generating reports related to analysis (e.g., regarding the cellular composition of a biological sample).

[0085] In some embodiments, the image processing module 530 includes one or more algorithms or models, such as those disclosed in U.S. Patent Application No. 17 / 371,746, the contents of which are incorporated herein by reference in their entirety. Examples of algorithms or models include, but are not limited to, identification algorithms for identifying sets of pixels in an image that correspond to individual components (e.g., cells) present in a biological sample; feature extraction algorithms for determining imaging features of individual components (e.g., cells) present in a biological sample based on the pixel values ​​of the identified sets of pixels; one or more differentiation models for different types of blood cells (e.g., different types of blood cells such as red blood cells, white blood cells, and platelets) based on the imaging features of individual components present in a biological sample; and cell counting algorithms for obtaining aggregate numbers for each type of cell present in a biological sample.

[0086] In some embodiments, the image processing module 530 also includes one or more mixers / unmixers, for example, algorithms configured to duplicate and mix a first signal and a second signal to generate a set of spectral spatial responses. In some embodiments, the first signal and the second signal represent the response of the sample to light at a first wavelength or wavelength range and at a second wavelength or wavelength range, respectively. In some embodiments, one or more mixers / unmixers include a non-temporary computer-readable medium for storing instructions, which, when executed by the processor, perform machine learning algorithms to count, size, and / or speciation of one or more cell types in the sample.

[0087] In various embodiments, one or more of the identified elements described above are stored in one or more of the aforementioned memory devices and correspond to a set of instructions for performing the above functions. The identified modules, data, or programs (e.g., sets of instructions) do not need to be implemented as separate software programs, procedures, datasets, or modules, and various subsets of these modules and data may be combined or otherwise rearranged in various embodiments. In some embodiments, non-persistent memory 511 optionally stores a subset of the identified modules and data structures described above. Furthermore, in some embodiments, the memory stores additional modules and data structures not described above. In some embodiments, one or more of the identified elements described above, other than those of computing device 501, are stored in the computing device and are addressable by computing device 501 so that all or part of such data can be retrieved as needed.

[0088] While computing device 501 is presented as a computer system, other topologies of computing device 501 are possible. For example, in some embodiments, computing device 501 can actually constitute several computer systems linked together in a network, or it can be a virtual machine or container in a cloud computing environment. Thus, Figure 5 shows an exemplary topology that is merely useful to illustrate the features of the embodiments of this disclosure in a manner easily understood by those skilled in the art.

[0089] In some embodiments, one or more components of the computing device 501 may be embedded in or integrated with the imaging device. For example, in one embodiment, a processor is integrated with the imaging device to process light detected by one or more detection assemblies of the imaging device, or captured images.

[0090] In some embodiments, the system 500 is configured to analyze biological samples such as blood samples, plasma samples, buffy coat samples, and urine samples. In some embodiments, the biological sample includes a blood cell sample containing one or more types of cells, e.g., red blood cells (RBCs), white blood cells (WBCs), and platelets. In some embodiments, the biological sample includes analytes present in urine, e.g., various types of crystals present in urine. In some embodiments, the sample is processed and tagged with a fluorescent marker of interest, such as a cell surface antigen or DNA / RNA stain. The sample is then mounted on an imaging device that captures one or more datasets of spectral, spatial, and / or temporal responses from the sample in response to irradiation and / or excitation. The data is analyzed, for example, on a computing device to provide information about the sample. In some embodiments, the system is configured to identify the total number of cells in the analyte, as well as their types, e.g., red blood cells (RBCs), platelets, white blood cells (WBCs), and different types thereof. In some embodiments, a combination of spatial, spectral, and / or temporal information about these cells enables rapid and reliable identification, counting, and speciation of different cell types within a sample. In some embodiments, the system is configured to provide identification of the types of crystals present in a urine sample.

[0091] In some embodiments, one or more light source assemblies of the imaging device 560 emit light at wavelengths independently selected from the group consisting of 405 nm, 460 nm, 470 nm, 520 nm, and 638 nm. In some embodiments, one or more light source assemblies include each light source configured to emit light at one of 405 nm, 460 nm, 470 nm, 520 nm, and 638 nm. For example, in some embodiments, light source assembly 140 includes at least one LED emitting light at 460 nm and at least one LED emitting light at 470 nm to excite DNA / RNA staining. In some embodiments, light source assembly 140 includes one or more LEDs emitting light at a single wavelength or a single wavelength range to excite DNA / RNA staining.

[0092] In some embodiments, the imaging device 560 is configured to acquire different types of images. For example, in some embodiments, the imaging device is configured to acquire fluorescence emission from the sample (e.g., fluorescence emission from analytes such as blood cells or urinary crystals in the sample), backscatter from the sample (e.g., backscatter from analytes such as blood cells or urinary crystals in the sample), transmission from the sample (e.g., transmission from analytes such as blood cells or urinary crystals in the sample, from which absorption and / or attenuation values ​​can be derived), bright-field imaging of the sample, and so on. In some embodiments, the absorption value derived from the transmission image is used to determine the volume of the analyte (e.g., cell volume or urinary crystal volume).

[0093] Referring to Figure 6, a flowchart is shown illustrating exemplary methods 600 for imaging and / or analyzing biological samples according to some exemplary embodiments of the present disclosure. In the flowchart, preferred parts of the method are shown in solid boxes, and additional, optional, or alternative parts of the method are shown in dashed boxes. Note that the processes disclosed herein and illustrated in the flowchart may, but are not required to be, performed without omission or in the order they are presented.

[0094] Method 600 includes, in step 610, placing a sample at a working distance from the lens assembly of an imaging device (e.g., imaging device 100, 200, 300, or 400). In some embodiments, the sample is a blood sample, plasma sample, buffy coat sample, urine sample, etc. The short working distance of the imaging device's lens assembly is less than 5 mm, less than 4 mm, less than 3 mm, or less than 2 mm. In addition to the lens assembly, the imaging device includes one or more light source assemblies and one or more detection assemblies. The lens assembly, at least one light source assembly, and at least one detection assembly are arranged to form an epitaxial configuration. In some embodiments, placing a sample at a working distance includes adjusting the focus and / or FOV of the imaging device. In some embodiments, once the sample is placed at a working distance from the imaging device's lens assembly, the focus and / or FOV of the imaging device is automatically adjusted.

[0095] Method 600 also includes, in step 620, activating at least one light source assembly of the imaging device to supply irradiation and / or excitation light to the sample. In one embodiment, the method activates one light source assembly, or individual light sources of a light source assembly, at one time. In another embodiment, the method activates two or more light source assemblies, or two or more individual light sources, simultaneously. In some embodiments, when activated, the light source assembly emits light containing excitation light in one or more excitation wavelength ranges to excite one or more components of the sample. For example, in some embodiments, the activated light source assembly emits light containing excitation light at a specific wavelength(s) to excite DNA / RNA staining. When excited, one or more components of the sample emit synchrotron radiation in one or more emission wavelength ranges. For example, in some embodiments, a DNA / RNA stain emits green and red light when excited.

[0096] Method 600 further comprises detecting light from a sample using one or more detection assemblies of an imaging device. Depending on the configuration of the imaging device (e.g., the arrangement of one or more light source assemblies relative to the sample, the spectrum of light from one or more light source assemblies), the detection procedure (e.g., which light source assemblies are activated), and / or other factors, the light from the sample may include light emitted from the sample, light scattered by the sample, light reflected by the sample, light transmitted through the sample, or any combination thereof. Depending on the configuration of the imaging device (e.g., the presence or absence of other optical systems such as filters, splitters, absorbers, etc.), and / or other factors, the light reaching one or more detection assemblies may be substantially the same as or different from the light from the sample. In one embodiment, the light reaching one or more detection assemblies is emitted light (e.g., fluorescence emission) at a single wavelength or a single wavelength range. In another embodiment, the light reaching one or more detection assemblies is emitted light at multiple (e.g., two, three, four, or more) wavelengths or wavelength ranges. In yet another embodiment, the light reaching one or more detection assemblies is light scattered by the sample, light reflected by the sample, and / or light transmitted through the sample. In yet another embodiment, the light reaching one or more detection assemblies is a combination of synchrotron radiation and light scattered by the sample, light reflected by the sample, and / or light transmitted through the sample.

[0097] One or more detection assemblies detect light from a sample (e.g., capture an image of an area of ​​the sample). In some embodiments, additional images are captured at different wavelengths by supplying a biological sample with illumination and / or excitation light of different wavelengths and detecting light from the sample in response to the illumination and / or excitation light of different wavelengths. In some embodiments, the illumination and / or excitation light of different wavelengths are supplied to the sample continuously, and the detection of light from the sample is performed continuously. Alternatively, in some embodiments, the illumination and / or excitation light of different wavelengths are supplied to the sample simultaneously, and the detection of light from the sample is performed simultaneously.

[0098] In some embodiments, Method 600 also includes, for example, processing detected light (e.g., a captured image) in a computing device 501 to generate one or more sets of spatial responses, temporal responses, and / or spectral responses in order to provide information about the sample. For example, in one embodiment, one or more detection assemblies receive light emitted from the sample. The method processes the detected synchrotron radiation to generate one or more sets of spectral responses and / or spatial responses, each set representing the spatial distribution of a corresponding component (e.g., a particular type of cell) within one or more components across an area of ​​the sample.

[0099] In some embodiments, the biological sample is a sample of blood cells containing red blood cells (RBCs), white blood cells (WBCs), and platelets. The method processes detected light (e.g., a captured image) to identify RBCs, WBCs, platelets, and their different types, and / or provides a whole blood count. In some embodiments, the biological sample includes analytes present in urine, e.g., various types of crystals present in urine. Method 600 processes detected light to identify the types of crystals present in the urine sample.

[0100] Terminology and References The terms used herein are for the purpose of describing specific embodiments and are not intended to limit the scope of the claims. As used in the description of embodiments and in the appended claims, the singular forms "a," "an," and "the" are intended to include the plural form unless otherwise explicitly indicated in the context. Terms such as "left" or "right," "top" or "bottom," "bottom" or "top," "internal" or "external" will be understood to be used to describe features of exemplary embodiments with reference to the arrangement of features as shown in the figures. Terms such as "first," "second," etc., may be used herein to describe various elements, but it will be understood that these elements should not be limited to these terms. These terms are used merely to distinguish one element from another. For example, as long as "first element" and "second element" are consistently interchangeable, the first element may be called the second element, and similarly, the second element may be called the first element, without changing the meaning of the description.

[0101] As used herein, the terms “and / or” refer to and encompass any possible combination of one or more of the related items listed. It will be further understood that, as used herein, the terms “comprise” and / or “comprising” specify the presence of the described features, components, steps, actions, elements, and / or components, but do not exclude the presence or addition of one or more other features, components, steps, actions, elements, components, and / or groups thereof.

[0102] As used herein, the terms “about” or “approximately” can mean within a tolerance range for a particular value as determined by those skilled in the art, which may in part depend on how that value is measured or determined, for example, the limits of the measuring system. For example, “about” can mean within or above one standard deviation, according to convention in the art. “About” can mean within ±20%, ±10%, ±5%, or ±1% of a given value. Where a particular value is described in this application and claims, unless otherwise specified, the term “about” means within a tolerance range for that particular value. The term “about” can have meanings as generally understood by those skilled in the art. The term “about” can mean ±10%. The term “about” can mean ±5%.

[0103] All references cited herein are incorporated herein by reference in whole, regardless of their purpose, to the same extent that each individual publication or patent or patent application is specifically and individually indicated to be incorporated by reference in whole, regardless of its purpose.

Claims

1. A lens assembly comprising a composite lens configured to optically communicate with a sample chamber in a detection area, wherein the lens assembly has (i) an optical path of less than about 15 mm, (ii) a working distance of about 5 mm or less, (iii) a weight of about 50 grams or less, or any combination thereof. A first light source assembly comprising a first light source configured to emit light into the sample chamber in the detection area, wherein the first light source assembly is positioned on the same side of the sample chamber as the lens assembly, and One or more detection assemblies that are optically in communication with the sample chamber in the detection area, wherein the one or more detection assemblies are configured to detect light from the sample chamber passing through the lens assembly in the detection area, An imaging device including, The imaging device is configured to define a first optical path through which light emitted from the first light source passes through the lens assembly and is guided in the detection area to the sample chamber. The imaging device is configured such that light returning from the sample chamber passes through the lens assembly in the detection area on the sample stage assembly, and defines a second optical path in which at least a portion of the light returning from the sample chamber in the detection area is guided to one or more detection assemblies. The aforementioned imaging device.

2. The imaging apparatus according to claim 1, wherein the lens assembly has more than three optical elements.

3. The first optical element among the three or more optical elements closest to the object-side plane of the lens assembly has a first effective aperture diameter and is at a first distance from the object-side plane of the lens assembly. The second optical element among the three or more optical elements that is furthest from the object-side plane of the lens assembly has a second effective aperture diameter and is at a second distance from the object-side plane of the lens assembly. The imaging apparatus according to claim 1 or 2, wherein the first effective aperture diameter is greater than or equal to the second effective aperture diameter, and the first distance is 0.4 times or more the second distance.

4. The imaging apparatus according to claim 3, wherein the first distance defines the working distance of the lens assembly.

5. The imaging apparatus according to claim 3 or 4, wherein the first optical element of the three or more optical elements has variable optical power radially from the optical axis of the first optical element or the lens assembly.

6. The imaging apparatus according to claim 5, wherein the variable optical power of the first optical element is to reduce aberrations along the optical axis, or both, enabling a wider field of view.

7. The imaging apparatus according to any one of the prior claims, further comprising a sample stage assembly configured to position the sample chamber in the detection area.

8. The imaging apparatus according to claim 7, wherein the sample stage assembly is configured to move the sample chamber relative to the lens assembly.

9. The imaging apparatus according to claim 7 or 8, wherein the sample stage assembly includes a rotating spindle configured to rotate the sample chamber with respect to the optical axis of the lens assembly in order to enable selection of a rotating field of view.

10. The imaging apparatus according to any one of claims 7 to 9, wherein the sample stage assembly is configured to move the sample chamber away from the detection area and to place a different sample chamber in the detection area.

11. The imaging apparatus according to any one of the prior claims, wherein the lens assembly has a rear focal plane of 8 mm or less, 7.5 mm or less, 7 mm or less, 6.5 mm or less, 6 mm or less, 5.5 mm or less, or 5 mm or less.

12. The imaging apparatus according to any one of the prior claims, wherein the composite lens includes a plurality of lenses packaged in a plastic lens barrel.

13. The aforementioned plastic lens barrel surrounds the optical axis of the composite lens and has an object side and an image side. The apparatus according to claim 12, wherein the composite lens comprises a first imaging lens having a relatively short rear focal length on the object side and a second imaging lens having a relatively long rear focal length.

14. The imaging apparatus according to claim 13, wherein the composite lens produces a magnification of 1x, 2x, 5x, 10x, 15x, 20x, or greater.

15. The imaging apparatus according to any one of the prior claims, wherein the composite lens includes 2 to 10 lenses.

16. Further comprising a first dichroic mirror, the first dichroic mirror is (i) Having a passband that includes either (a) the excitation wavelength band emitted from the first light source, or (b) one or more emission wavelength bands, (ii) Having a stopband that includes (a) the excitation wavelength band emitted from the first light source, or (b) the other of the one or more emission wavelength bands, (iii) Arranged in the first optical path between the lens assembly and the first light source assembly, (iv) The imaging device according to any one prior claim, which is positioned in the second optical path between the lens assembly and the one or more detection assemblies.

17. The present invention further includes a second dichroic mirror positioned in the second optical path between the first dichroic mirror and the one or more detection assemblies, (i) The second dichroic mirror has a passband that includes a first portion of the one or more emission wavelength bands, (ii) The second dichroic mirror has a stopband that includes a second portion of the one or more emission wavelength bands, (iii) The one or more detection assemblies include a first detection assembly comprising a first two-dimensional photodetector configured to acquire an image in the wavelength band of the first portion of the one or more emission wavelength bands, (iv) The imaging apparatus according to claim 16, wherein the one or more detection assemblies include a second detection assembly comprising a second two-dimensional photodetector configured to acquire an image in the wavelength band of the second portion of the one or more emission wavelength bands.

18. The first detection assembly includes a first tube lens positioned between the second dichroic mirror and the first two-dimensional photodetector. The imaging apparatus according to claim 17, wherein the second detection assembly includes a second tube lens positioned between the second dichroic mirror and the second two-dimensional photodetector.

19. At least one splitter, and It further includes multiple filters, The one or more detection assemblies include a plurality of detection assemblies, The at least one splitter is positioned in the second optical path between the first dichroic mirror and the plurality of detection assemblies and is configured to divide the one or more emission wavelength bands into a plurality of parts. Each of the plurality of filters is positioned between the at least one splitter and the corresponding detection assembly in the plurality of detection assemblies, and has a passband for each of the plurality of passbands, and filters the corresponding portion of the plurality of portions of the one or more emission wavelength bands. The imaging apparatus according to claim 16, wherein the corresponding detection assembly in the plurality of detection assemblies includes a two-dimensional photodetector configured to acquire an image of the corresponding portion in the plurality of portions of the one or more emission wavelength bands after passing through the respective filters of the plurality of filters.

20. The imaging apparatus according to any one of the prior claims, further comprising a voice coil actuator coupled to or integrated with the lens assembly, configured to adjust the focus of the composite lens.

21. The imaging apparatus according to claim 20, wherein the voice coil actuator is smaller than 20 mm in width, 20 mm in length, and 20 mm in height.

22. The imaging apparatus according to claim 20, wherein the focal point of the composite lens of the lens assembly is adjustable within a range of approximately ±2 mm, approximately ±1.5 mm, approximately ±1.0 mm, or approximately ±0.75 mm or less.

23. Further including a second light source assembly, The second light source assembly is positioned on the opposite side of the sample stage assembly as the lens assembly. The second light source assembly is configured to emit light into the sample chamber in the detection area, The imaging device according to any one of the prior claims, wherein the imaging device is configured such that light emitted from the second light source assembly passes through the sample chamber and the lens assembly in the detection area, and defines a third optical path in which at least a portion of the light that has passed through the sample chamber in the detection area is guided to one or more detection assemblies.

24. The imaging apparatus according to claim 23, wherein the second light source assembly includes a bright-field light source, a dark-field light source, a side-scatter light source, or any combination thereof.

25. The bright-field light source is directed approximately 45 degrees relative to the irradiation area of ​​the sample. ° ~about 90 ° The imaging apparatus according to claim 24, wherein the sample chamber is irradiated at the angle.

26. The dark-field light source is approximately 0 in relation to the irradiation area of ​​the sample. ° ~about 45 ° The imaging apparatus according to claim 24 or 25, wherein the sample chamber is irradiated at the angle.

27. The imaging apparatus according to any one of claims 24 to 26, wherein the second light source assembly further includes a diffuser for softening or diffusing light generated by the bright-field light source, the dark-field light source, or the side-scatter light source.

28. The imaging apparatus according to any one of claims 23 to 27, wherein the control unit is configured to selectively operate the first light source assembly, the second light source assembly, any individual light source in the first light source assembly, any individual light source in the second light source assembly, or any combination thereof.

29. The imaging apparatus according to any one of the prior claims, wherein the sample chamber is part of a centrifugal microfluidic biodisk, part of a capillary apparatus, or part of a flow cell apparatus.

30. The imaging apparatus according to claim 29, wherein the centrifugal microfluidic biodisk includes an elongated reservoir radially positioned at a first distance from the axis of rotation of the centrifugal microfluidic biodisk.

31. The imaging apparatus according to any one of the prior claims, wherein the composite lens is a general-purpose lens arranged in reverse so that the sample chamber is positioned at the rear focal plane of the general-purpose lens.

32. The imaging device according to any one of the claims, wherein the dimensions of the imaging device are less than 25 centimeters (cm) in length, less than 25 cm in width, and less than 25 cm in height.

33. The imaging apparatus according to any one of the prior claims, wherein the first light source assembly is configured to emit light in a single narrow wavelength band.

34. The imaging apparatus according to claim 33, wherein the full width at half maximum (FWHM) of the single narrow wavelength band is 50 nm or less, 25 nm or less, 10 nm or less, or 5 nm or less.

35. The imaging apparatus according to any one of the prior claims, wherein the first light source assembly includes one or more light-emitting diodes (LEDs).

36. The imaging apparatus according to claim 35, wherein the one or more LEDs include a first LED configured to emit light in a first narrow wavelength band and a second LED configured to emit light in a second narrow wavelength band.

37. The imaging apparatus according to any one of the prior claims, wherein the first light source is a single LED.

38. The imaging apparatus according to any one of the prior claims, wherein the overall length of the lens assembly is approximately 5 mm to approximately 20 mm, and the diagonal dimension of the lens assembly on the object-side plane is approximately 3 mm to 20 mm.

39. The imaging device according to claim 38, wherein the overall length is approximately 10 mm, and the diagonal dimensions are approximately 5.0 mm, approximately 5.8 mm, approximately 7.0 mm, approximately 8.0 mm, approximately 9.0 mm, or approximately 12.0 mm.

40. The imaging device according to claim 38, wherein the overall length is approximately 7 mm, and the diagonal dimensions are approximately 5.0 mm, approximately 5.8 mm, approximately 7.0 mm, approximately 8.0 mm, approximately 9.0 mm, or approximately 12.0 mm.

41. An imaging device according to any one of the prior claims, configured to capture one or more images of a sample in the sample chamber in the detection area, A control unit configured to communicate with the imaging device via wired or wireless means and to control the imaging device, and A computing device that communicates with the control unit by wire or wireless means and processes one or more captured images of the sample. A system that includes this.

42. The system according to claim 41, wherein the control unit is a standalone unit.

43. The system according to claim 41, wherein the control unit is embedded in or integrated with the imaging device.

44. The system according to claim 41, wherein the control unit is embedded in or integrated with the computing device.

45. The sample chamber containing the sample is placed at the working distance from the lens assembly of the imaging apparatus according to any one of claims 1 to 40, Activating the first light source assembly to supply excitation light to the sample, The one or more detection assemblies detect the synchrotron radiation emitted from the sample, Methods that include...

46. The method according to claim 45, further comprising processing the detected synchrotron radiation with a computing device to generate one or more sets of spectral responses and / or spatial responses, each set representing the spatial distribution of corresponding components within one or more components over an area of ​​the sample.

47. The method according to claim 45 or 46, wherein the sample is blood or urine.

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