Measurement using rapid temperature changes

Abstract

The present invention provides devices, systems and methods for rapidly and easily promoting reactions (such as but not limited to PCR) during thermal cycling or temperature changes of samples.

Description

[0001] This application is a divisional application of the Chinese national phase application with application number 2018800249487 and filing date 2018-02-15.

[0002] This application claims the benefit of USPA (hereinafter "USPA") No. 62 / 460,088 filed on February 16, 2017 (053PRV), USPA No. 62 / 460,083 filed on February 16, 2017 (035PRV3), USPA No. 62 / 460,076 filed on February 16, 2017 (040PRV2), USPA No. 62 / 460,075 filed on February 16, 2017 (026PRV3), USPA No. 62 / 460,076 filed on February 16, 2017 (036PRV3), USPA No. 62 / 460,077 filed on February 16, 2017 (046PRV3), USPA No. 62 / 460,078 filed on February 16, 2017 (081PRV4), USPA No. 62 / 460,079 filed on February 16, 2017 (091PRV5), USPA No. 62 / 460,080 filed on February 16, 2017 (083PRV6), USPA No. 62 / 460,081 filed on February 16, 2017 (093PRV7), USPA No. 62 / 460,083 filed on February 16, 2017 (083PRV8), USPA No. 62 / 460,084 filed on February 16, 2017 (083PRV9), USPA No. 62 / 460,085 filed on February 16, 2017 (083PRV1 USPA No. 62 / 460,069 filed on February 16, 2017 (052PRV2), USPA No. 62 / 460,062 filed on February 16, 2017 (045PRV2), USPA No. 62 / 460,047 filed on February 16, 2017 (029PRV2), USPA No. 62 / 459,972 filed on February 16, 2017 (002PRV2), USPA No. 62 / 460,063 filed on February 16, 2017 (050PRV2), USPA No. 62 / 460,064 filed on February 16, 2017 (045PRV2), USPA No. 62 / 460,065 filed on February 16, 2017 (029PRV2), USPA No. 62 / 459,976 filed on February 16, 2017 (002PRV2), USPA No. 62 / 460,066 filed on February 16, 2017 (054PRV2), USPA No. 62 / 460,067 filed on February 16, 2017 (054PRV2), USPA No. 62 / 460,068 filed on February 16, 2017 (055PRV2), USPA No. 62 / 460,0 No. 2 / 459,920, USPA No. 62 / 459,602 filed on February 15, 2017 (052PRV), USPA No. 62 / 459,598 filed on February 15, 2017 (035PRV2), USPA No. 62 / 459,577 filed on February 15, 2017 (051PRV), USPA No. 62 / 459,554 filed on February 15, 2017 (026PRV2), USPA No. 62 / 459,603 filed on February 15, 2017 (035PRV2), USPA No. 62 / 459,610 filed on February 15, 2017 (036PRV2), USPA No. 62 / 459,636 filed on February 15, 2017 (036PRV2), USPA No. 62 / 459,643 filed on February 15, 2017 (036PRV2), USPA No. 62 / 459,650 filed on February 15, 2017 (036PRV2), USPA No. 62 / 459,661 filed on February 15, 2017 ( 014PRV2), USPA No. 62 / 459,496 filed on February 15, 2017 (033PRV2), USPA No. 62 / 459,337 filed on February 15, 2017 (033PRV2), USPA No. 62 / 459,303 filed on February 15, 2017 (027PRV2), USPA No. 62 / 459,267 filed on February 15, 2017 (017PRV2), USPA No. 62 / 459,496 filed on February 15, 2017 (014 ...No. 232, PCT Application No. PCT / US18 / 18108 filed on February 14, 2018 (18F14), PCT Application No. PCT / US18 / 18007 filed on February 13, 2018 (18F11B), PCT Application No. PCT / US18 / 17716 filed on February 9, 2018 (18F08), PCT Application No. PCT / US18 / 17713 filed on February 9, 2018 (18F07), PCT Application No. PCT / US18 / 17712 filed on February 9, 2018 (18F16), PCT Application No. PCT / US18 / 17504 filed on February 8, 2018 (18F02), PCT Application No. PCT / US18 / 17506 filed on February 8, 2018 PCT Application No. PCT / US18 / 17499 filed on February 8, 2018 (18F17), PCT Application No. PCT / US18 / 17489 filed on February 8, 2018 (18F18), PCT Application No. PCT / US18 / 17492 filed on February 8, 2018 (18F10), PCT Application No. PCT / US18 / 17494 filed on February 8, 2018 (18F02), PCT Application No. PCT / US18 / 17502 filed on February 8, 2018 (18F09), and PCT Application No. PCT / US18 / 17307 filed on February 7, 2018 (18F15A), each of which is incorporated herein by reference in its entirety for all purposes. Background Art

[0003] In some chemical, biological and / or medical assays, repeated thermal cycles and rapid and accurate temperature control are required. A specific example is the polymerase chain reaction (PCR), which is used to amplify predetermined nucleotides (e.g., DNA) in one or more samples. In PCR, samples are repeatedly heated and cooled to a specific temperature following a predetermined thermal control cycle. In some cases, it is necessary to be able to quickly and evenly change the temperature of the sample. Summary of the invention

[0004] The following brief summary is not intended to include all features and aspects of the present invention. The present invention provides devices, systems and methods for rapid sample thermal cycling changes to promote reactions such as but not limited to PCR. One aspect of the present invention is to use two movable thin plates to press a liquid sample into a uniform thin layer.

[0005] Another aspect of the present invention is to accelerate the sample temperature ramp rate, QMAX measurement of a plate down to 100 μm (micrometers) thick or less.

[0006] Another aspect of the present invention is to provide solutions to the problems associated with very thin plates in QMAX cards under rapid temperature changes or cycles. Problems include large deformations of the plates, sample thickness variations, bubble formation, etc., any of which can cause PRC or isothermal nucleic acid amplification to fail. Therefore, it is very important according to the present invention that one of the solutions involves the use of a fixture with a specific design to hold the two plates together during the temperature change. Of course, the fixture of the present invention can be used for QMAX cards that do not require changes in sample temperature.

[0007] The present invention provides an apparatus and method for rapidly changing the temperature of a sample by forming a uniform ultra-thin layer over an area (or an area of ​​interest), a sample holder with low heat absorption and low heat capacity, and an area heater element.

[0010] The present invention provides devices and methods for rapid thermal cycling changes, and the devices and methods disclosed herein are suitable for facilitating reactions such as, but not limited to, PCR. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Those skilled in the art will appreciate that the drawings described below are for illustrative purposes only. The drawings are not intended to limit the scope of the present invention in any way. In some cases, the drawings are not drawn completely to scale. In the figures where experimental data points are given, the lines connecting the data points are only used to guide the observed data and have no other meaning.

[0012] Figure 1 Perspective views and cross-sectional views of an embodiment of the device of the present invention are shown; Figure (A) shows an embodiment of the device in an open structure; Figure (B) shows an embodiment of the device when the sample unit is in a closed structure, wherein the temperature of the sample compressed into a thin layer between two plates is rapidly changed by a radiation source positioned to project electromagnetic waves onto the sample.

[0013] Figure 2 Perspective views and cross-sectional views of an embodiment of the system of the present invention are shown; Figure (A) shows a perspective view of the system when the device (sample unit of the system) is in an open structure; Figure (B) shows a cross-sectional view of the system when the sample unit is in a closed structure.

[0014] Figure 3 A cross-sectional view of an embodiment of the system of the present invention is shown, illustrating the system and showing additional elements that facilitate temperature change and control.

[0015] Figure 4 A perspective view of another embodiment of the present invention is shown, wherein there are multiple sample contact areas on the plate, allowing multiple samples to be processed and analyzed.

[0016] Figure 5A cross-sectional view of an exemplary embodiment of the present invention is shown, illustrating how a sample is added and compressed.

[0017] Figure 6 A cross-sectional view of an exemplary embodiment of the present invention is shown, illustrating the PCR process.

[0018] Detailed Description of Exemplary Embodiments

[0019] The following detailed description illustrates some embodiments of the present invention by way of example and not limitation. The section headings and any subheadings used herein are for organizational purposes only and should not be interpreted as limiting the subject matter described in any way. The content under the section headings and / or subtitles is not limited to the section headings and / or subtitles, but applies to the entire description of the present invention.

[0020] The citation of any publication is for its disclosure prior to the filing date and should not be construed as an admission that the present claims are not entitled to antedate such publication by virtue of prior invention. Further, the publication dates provided may be different from the actual publication dates which may need to be independently confirmed.

[0021] It should be noted that the drawings are not intended to show elements in strict proportion. For the sake of clarity, some elements are enlarged when shown in the drawings. The dimensions of the elements should be depicted according to the description provided herein and incorporated herein by reference.

[0022] QMAX Device

[0023] The terms "CROF card (or card)", "COF card", "QMAX card", "Q card", "CROF device", "COF device", "QMAX device", "CROF board", "COF board" and "QMAX board" are interchangeable, except that in some embodiments, the COF card does not include a spacer; and these terms refer to a device that includes a first board and a second board that are movable relative to each other into different configurations (including an open configuration and a closed configuration), and the device includes a spacer that adjusts the spacing between the boards (except for some embodiments of COF). The term "X board" refers to one of the two boards in a CROF card, wherein the spacer is fixed to the board. Further description of COF cards, CROF cards and X boards are described in provisional application serial number 62 / 456065 filed on February 7, 2017, the entire contents of which are incorporated herein for all purposes.

[0024] Figure 1A perspective view and a cross-sectional view of an embodiment of the device of the present invention are shown. Figure (A) shows a device (also referred to as a "sample unit" of the system) 100 in an open configuration. As shown in Figure (A), the sample unit 100 comprises a first plate 10, a second plate 20 and a spacing mechanism (not shown). The first plate 10 and the second plate 20 respectively comprise an outer surface (respectively 11 and 21) and an inner surface (respectively 12 and 22). Each inner surface has a sample contact area (not shown) for contacting a fluid sample to be processed and / or analyzed by the device.

[0025] The first plate 10 and the second plate 20 can be moved relative to each other into different configurations. One of these configurations is an open configuration, in which, for example Figure 1 As shown in FIG. 1A , the first plate 10 and the second plate 20 are partially or completely separated, and the spacing between the first plate 10 and the second plate 20 (i.e., the distance between the first plate inner surface 11 and the second plate inner surface 21) is not adjusted by the spacing mechanism. The open configuration allows the sample to be deposited on the first plate, the second plate, or both in the sample contact area.

[0026] like Figure 1 As shown in FIG. (A), the first plate 10 also includes a radiation absorbing layer 112 located in the sample contact area. It is also possible that the second plate 20 optionally or additionally includes a radiation absorbing layer 112. In some embodiments, the radiation absorbing layer 112 is configured to effectively absorb radiation (e.g., electromagnetic waves) emitted thereon. The absorption percentage is 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 99% or more, 100% or less, 85% or less, 75% or less, 65% or less, or 55% or less, or in a range between any two values. The radiation absorbing layer 112 is also configured to convert at least most of the absorbed radiation energy into heat (thermal energy). For example, the radiation absorbing layer 112 is configured to emit radiation in the form of heat after absorbing energy from electromagnetic waves. As used herein, the terms "majority" or "substantially" refer to a percentage of 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, 99% or more, 99% or more, or 99.9% or more.

[0027] In some embodiments, the radiation absorbing layer 112 comprises a material / structure such as, but not limited to, a metal plasma surface, a metamaterial, black silicon, graphite, carbon nanotubes, silicon interlayers, graphene, superlattices, plasma materials, any material / structure that can effectively absorb electromagnetic waves and convert the absorbed energy into heat energy, and any combination thereof. In some embodiments, the radiation absorbing layer 112 comprises carbon nanotubes.

[0028] In some embodiments, the radiation absorbing layer comprises a point-coupled point-pole antenna (D2PA) array, such as, but not limited to, the D2PA arrays described in U.S. Provisional Patent Application No. 61 / 347,178 filed on May 21, 2010, U.S. Provisional Patent Application No. 61 / 622,226 filed on April 10, 2012, U.S. Provisional Patent Application No. 61 / 801,424 filed on March 15, 2013, U.S. Provisional Patent Application No. 61 / 801,096 filed on March 15, 2013, U.S. Provisional Patent Application No. 61 / 801,933 filed on March 15, 2013, and U.S. Provisional Patent Application No. 61 / 801,943 filed on March 1, 2013. U.S. Provisional Patent Application No. 61 / 794,317 filed on December 5, 2014, U.S. Provisional Patent Application No. 62 / 090,299 filed on December 10, 2014, U.S. Provisional Patent Application No. 61 / 708,314 filed on October 1, 2012, PCT Application No. PCT / US2011 / 037455 filed on May 20, 2011, PCT Application No. PCT / US2013 / 032347 filed on March 15, 2013, PCT Application No. PCT / US2014 / 029979 filed on March 14, 2014, PCT Application No. PCT / US2014 / 028417, PCT Application No. PCT / US2014 / 030108 filed on March 16, 2014, PCT Application No. PCT / US2013 / 062923 filed on October 1, 2013, U.S. Patent Application No. 13 / 699,270 filed on June 13, 2013, U.S. Patent Application No. 14 / 459,239 filed on August 13, 2014, U.S. Patent Application No. 14 / 871,678 filed on September 30, 2015, U.S. Patent Application No. 13 / 838,600 filed on March 15, 2013, and U.S. Patent Application No. 13 / 838,600 filed on August 13, 2014 U.S. patent application number 14 / 459,251, U.S. patent application number 14 / 668,750 filed on March 25, 2015, U.S. patent application number 14 / 775,634 filed on September 11, 2015, U.S. patent application number 14 / 775,638 filed on September 11, 2015, U.S. patent application number 14 / 852,412 filed on March 16, 2014, U.S. patent application number 14 / 964,394 filed on December 9, 2015, and U.S. patent application number 14 / 431,266 filed on October 5, 2015, the entire disclosures of which are incorporated herein by reference for all purposes.

[0029] Figure 1FIG. (B) shows a perspective view and a cross-sectional view of the sample unit 100 in a closed configuration. The cross-sectional view shows a portion of the device without showing the entirety of the sample unit 100 or the spacing mechanism. As shown in FIG. (B), the sample unit 100 includes a first plate 10, a second plate 20, and a spacing mechanism (not shown).

[0030] exist Figure 1 In the figure (B), the first plate 10 and the second plate 20 are in a closed configuration. In the closed configuration, the inner surfaces of the two plates 11 and 21 face each other, and the spacing between the two plates 102 is adjusted by the spacing mechanism. Therefore, as shown in the figure, the two plates press the fluid sample 90 on one or both of the deposition plates into a layer, and the thickness of the layer is adjusted by the spacing mechanism (not shown).

[0031] NN1. An apparatus for rapidly changing the temperature of a thin fluid sample layer, comprising:

[0032] A first plate and a second plate, wherein:

[0033] i. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample; and

[0034] ii. The plate has a configuration for rapidly changing the temperature of the sample, wherein:

[0035] a. The sample contact areas face each other and are substantially parallel,

[0036] b. The average spacing between contact areas is equal to or less than 200 microns,

[0037] c. the two plates condition (or define) at least a portion of the sample into a layer of very uniform thickness and substantially stationary relative to the plates,

[0038] d. a radiation absorbing layer is located adjacent to at least a portion of the sample of uniform thickness,

[0039] e. At least a portion of the sample and the region of the radiation absorbing layer are of substantially greater than uniform thickness.

[0040] In some embodiments of the invention, there is a spacer between the two plates.

[0041] In some embodiments, during heating, there is an "anti-evaporation ring" outside the liquid area (eg, sample area) that prevents or reduces vapor of the liquid from escaping the card.

[0042] In some embodiments, there is a clamp on the outside of the QMAX card to secure the QMAX card in its closed configuration during heating.

[0043] In some embodiments, in order to achieve rapid and uniform thermal changes in the sample, the sample is pressed into a thin layer. The thickness of the layer is 500 μm or less, 200 μm or less, 100 μm or less, 50 μm or less, 20 μm or less, 10 μm or less, 5 μm or less, 2 μm or less, 1 μm or less, 500 nm or more, 1.5 μm or more, 2.5 μm or more, 7.5 μm or more, 15 μm or more, 30 μm or more, 75 μm or more, 150 μm or more, or 250 μm or more. The thin thickness of the sample layer allows the reagent to diffuse faster and / or the heat to conduct faster. In some embodiments, the two plates are pressed with an imprecise pressure that is neither set to an exact level nor substantially uniform. In certain embodiments, the two plates are pressed directly with a human hand.

[0044] In some embodiments, the QMAX card, including the plates and spacers, is made of a material with low thermal conductivity to reduce heat absorption by the card itself.

[0045] In some embodiments, there is a clamp on the outside of the QMAX card to secure the QMAX card in its closed configuration during heating.

[0046] The clamp is also made of a material with low thermal conductivity to reduce heat absorption by the card itself.

[0047] The term "cover" refers to the area on the board that is in thermally conductive contact with the clamp. For example, the clamp covering some surface of the QMAX card in a closed configuration means that the clamp has thermally conductive contact with a portion of the board surface of the QMAX card.

[0048] The term "sealing" by the clamp means that the clamp, in a closed configuration of the plate, prevents or reduces the flow of at least a portion of the sample from one location of the plate to another location of the plate.

[0049] For example, the clamp may be configured to seal a portion of a sample or the entire sample.

[0050] The term "clamp" refers to a device comprising two elements that insert a compressive force to hold a third element or elements together. For example, a clamp holds two plates together.

[0051] Wherein these materials include but are not limited to polymers (such as plastics) or amorphous organic materials. Polymeric materials include but are not limited to acrylate polymers, vinyl polymers, olefin polymers, cellulosic polymers, non-cellulosic polymers, polyester polymers, nylon, cyclic olefin copolymers (COC), poly(methyl methacrylate) (PMMA), polycarbonate (PC), cyclic olefin polymers (COP), liquid crystal polymers (LCP), polyamide (PA), polyethylene (PE), polyimide (PI), polypropylene (PP), polyphenylene ether (PPE), polystyrene (PS), polyoxymethylene (POM), polyetheretherketone (PEEK), polyethersulfone (PES), polyethylene phthalate (PET), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), polyvinylidene fluoride (PVDF), polybutylene terephthalate (PBT), fluorinated ethylene propylene (FEP), perfluoroalkoxyalkane (PFA), polydimethylsiloxane (PDMS), rubber or any combination thereof.

[0052] These materials include but are not limited to inorganic materials, including dielectric materials such as silicon oxide, porcelain, ceramics, mica, glass, and oxides of various metals.

[0053] These materials include but are not limited to inorganic compounds, including aluminum oxide, aluminum chloride, cadmium sulfide, gallium nitride, gold chloride, indium arsenide, lithium borohydride, silver bromide, sodium chloride, etc.

[0054] These materials include liquids, including but not limited to water, ethane, methane, oil, benzene, hexane, heptane, silicone oil, polychlorinated biphenyls, liquid air, liquid oxygen, liquid nitrogen, etc.

[0055] These materials include gases, including but not limited to air, argon, helium, nitrogen, oxygen, carbon dioxide, etc.

[0056] The material is a combination of the above materials.

[0057] In some embodiments, at least one of the plates has an average thickness of 1 to 1000 μm, 10 to 900 μm, 20 to 800 μm, 25 to 700 μm, 25 to 800 μm, 25 to 600 μm, 25 to 500 μm, 25 to 400 μm, 25 to 300 μm, 25 to 200 μm, 30 to 200 μm, 35 to 200 μm, 40 to 200 μm, 45 to 200 μm, or 50 to 200 μm.

[0058] In some embodiments, at least one of the plates has an average thickness in a range of 50 μm to 75 μm, 75 μm to 100 μm, 100 μm to 125 μm, 125 μm to 150 μm, 150 μm to 175 μm, or 175 μm to 200 μm.

[0059] In some embodiments, the average thickness of at least one of the plates is about 50 μm, about 75 μm, about 100 μm, about 125 μm, about 150 μm, about 175 μm, or about 200 μm.

[0060] The height of the spacer is selected by the desired adjustment spacing between the plates and / or the final sample thickness of the adjustment and the remaining sample thickness. The spacer height (predetermined spacer height), the spacing between the plates and / or the sample thickness is 3 nm or less, 10 nm or less, 50 nm or less, 100 nm or less, 200 nm or less, 500 nm or less, 800 nm or less, 1000 nm or less, 1 μm or less, 2 μm or less, 3 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, 30 μm or less, 50 μm or less, 100 μm or less, 150 μm or less, 200 μm or less, 300 μm or less, 500 μm or less, 800 μm or less, 1 mm or less, 2 mm or less, 4 mm or less, or in a range between any two of these values.

[0061] The spacer height, spacing between plates and / or sample thickness is from 1 nm to 100 nm in one preferred embodiment, from 100 nm to 500 nm in another preferred embodiment, from 500 nm to 1000 nm in a separate preferred embodiment, from 1 μm (i.e. 1000 nm) to 2 μm in another preferred embodiment, from 2 μm to 3 μm in a separate preferred embodiment, from 3 μm to 5 μm in another preferred embodiment, from 5 μm to 10 μm in a separate preferred embodiment, and from 10 μm to 50 μm in another preferred embodiment, and from 50 μm to 100 μm in a separate preferred embodiment.

[0062] In some embodiments, the QMAX device is fully transparent or partially transparent to reduce heat absorption by the card itself, wherein the transparency is higher than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% or any range between two values.

[0063] In some embodiments, the QMAX device is partially reflective to reduce heat absorption by the card itself, wherein the reflectivity of the surface is greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or a range between any two values.

[0064] In some embodiments, the QMAX and the fixture are coated with a thermal insulation layer to reduce the heat absorption of the card itself, wherein the thermal insulation layer comprises a material including the low thermal conductivity material described above.

[0065] In some embodiments, the clamp covers and seals all QMAX cards in a closed configuration.

[0066] In some embodiments, the fixture covers some surface of the QMAX card in the closed configuration.

[0067] In some embodiments, the fixture has a transparent window to allow light to enter the interior of the QMAX card and exit the QMAX card.

[0068] In some embodiments, the fixture is completely transparent to allow light to enter the interior of the QMAX card and exit the QMAX card.

[0069] The transparency of the fixture is greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or a range between any two of these values.

[0070] In some embodiments, air or liquid is present between the clamp in the closed configuration and the QMAX device.

[0071] The liquid includes but is not limited to water, ethane, methane, oil, benzene, hexane, heptane, silicone oil, polychlorinated biphenyls, liquid air, liquid oxygen, liquid nitrogen, etc.

[0072] The gas includes but is not limited to air, argon, helium, nitrogen, oxygen, carbon dioxide and the like.

[0073] In some embodiments, after closing the clamp, the pressure applied by the clamp on the surface of the QMAX card is 0.01 kg / cm 2 , 0.1kg / cm 2 , 0.5kg / cm 2 , 1kg / cm 2 , 2kg / cm 2 、kg / cm 2 , 5kg / cm 2 , 10kg / cm 2 , 20kg / cm 2 、30kg / cm 2 , 40kg / cm 2 , 50kg / cm 2 、60kg / cm 2 , 100kg / cm 2 、150kg / cm 2 , 200kg / cm 2 , or the range between any two values; and 0.1kg / cm 2 Up to 0.5kg / cm 2 , 0.5kg / cm 2 Up to 1kg / cm 2, 1kg / cm 2 Up to 5kg / cm 2 , 5kg / cm 2 Up to 10kg / cm 2 Preferred range of (pressure).

[0074] like Figure 1 As shown in the cross-sectional view of the device in , the radiation absorbing layer 112 spans the sample contact area. However, it should be noted that the lateral area of ​​the radiation absorbing layer may also occupy only a portion of the sample contact area, with a percentage of about 1% or more, 5% or more, 10% or more, 20% or more, 50% or more, 80% or more, 90% or more, 95% or more, 99% or more, 85% or less, 75% or less, 55% or less, 40% or less, 25% or less, 8% or less, 2.5% or less. In some embodiments, to promote temperature changes in the sample, in some embodiments, the lateral area of ​​the radiation absorbing layer is configured so that the sample 90 receives thermal radiation from the radiation absorbing layer 112, and the thermal radiation is substantially uniform across the lateral dimensions of the sample 90 over the sample contact area.

[0075] In some embodiments, the radiation absorbing area is 10%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100% of the total panel area, or a range between any two values.

[0076] In some embodiments, the thickness of radiation absorbing layer 112 is 10 nm or more, 20 nm or more, 50 nm or more, 100 nm or more, 200 nm or more, 500 nm or more, 1 μm or more, 2 μm or more, 5 μm or more, 10 μm or more, 20 μm or more, 50 μm or more, 100 μm or more, 75 μm or less, 40 μm or less, 15 μm or less, 7.5 μm or less, 4 μm or less, 1.5 μm or less, 750 nm or less, 400 nm or less, 150 nm or less, 75 nm or less, 40 nm or less, or 15 nm or less, or in a range between any two values. In some embodiments, the thickness of radiation absorbing layer 112 is 100 nm or less.

[0077] In some embodiments, the area of ​​the sample layer and the radiation absorbing layer 112 is substantially greater than the uniform thickness. Here, the term "substantially greater" means that the approximate diameter or diagonal distance of the sample layer and / or the radiation absorbing layer is at least 10 times, 15 times, 20 times, 25 times, 30 times, 35 times, 40 times, 45 times, 50 times, 55 times, 60 times, 65 times, 70 times, 75 times, 80 times, 85 times, 90 times, 95 times, 100 times, 150 times, 200 times, 250 times, 300 times, 400 times, 450 times, 500 times, 550 times, 600 times, 650 times, 700 times, 750 times, 800 times, 850 times, 900 times, 950 times, 1000 times, 1500 times, 2000 times, 2500 times, 3000 times, 4000 times, 4500 times, 5000 times, 6000 times, 6500 times, 7000 times, 7500 times, 8000 times, 8500 times, 9000 times, 9500 times, 10000 times, 15000 times, 20000 times, 25000 times, 30000 times, 40000 times, 45000 times, 50000 times, 50000 times, 60000 times, 65000 times, 70000 times, 75000 times, 80000 times, 85000 times, 90000 times, 95000 times, 100 times, 350 times, 400 times, 450 times, 500 times, 550 times, 600 times, 650 times, 700 times, 750 times, 800 times, 850 times, 900 times, 950 times, 1000 times, 1500 times, 2000 times, 2500 times, 3000 times, 3500 times, 4000 times, 4500 times, or 5000 times, or in a range between any two values.

[0078] Systems including QMAX devices

[0079] Figure 2 The system of the present invention is shown in perspective and cross-sectional views. As shown in (A) and (B), the system comprises a sample unit 100 and a thermal control unit 200; the sample unit 100 comprises a first plate 10, a second plate 20 and a spacing mechanism (not shown); the thermal control unit 200 comprises a radiation source 202 and a controller 204. Figure 2 Figures (A) and (B) show a perspective view and a cross-sectional view of the system when the sample unit 100 of the system is in a closed configuration.

[0080] like Figure 1 As shown in FIG. 2B , the thermal control unit 200 includes a radiation source 202 and a controller 204. In some embodiments, the thermal control unit 200 provides energy in the form of electromagnetic waves for changing the temperature of the sample.

[0081] See also Figure 2 (A) and (B), the radiation source 202 is configured to project an electromagnetic wave 210 to the radiation absorbing layer 112 of the sample unit 100, which is configured to absorb the electromagnetic wave 210 and convert most of the electromagnetic wave 210 into heat to generate thermal radiation, which increases the temperature of the portion of the sample 90 close to the radiation absorbing layer 112. In other words, the coupling of the radiation source 202 and the radiation absorbing layer 112 is configured to generate the thermal energy required to promote the temperature change of the sample 90.

[0082] In some embodiments, the radiation from the radiation source 202 comprises radio waves, microwaves, infrared waves, visible light, ultraviolet waves, X-rays, gamma rays, or thermal radiation, or any combination thereof. In some embodiments, the radiation absorbing layer 112 has a preferred light wavelength range, within which the radiation absorbing layer 112 has the highest absorption efficiency. In some embodiments, the radiation source 202 is configured to project electromagnetic waves within, overlapping with, or including the preferred wavelength range of the radiation absorbing layer 112. In other embodiments, the wavelength is appropriately designed to be away from the preferred wavelength of the radiation absorbing layer in order to promote temperature changes.

[0083] In some embodiments, radiation source 202 comprises a laser source that provides laser light within a narrow wavelength range. In other embodiments, radiation source 202 comprises an LED (light emitting diode) or multiple LEDs.

[0084] Reference Figure 2 In Figures (A) and (B), the controller 204 is configured to control the electromagnetic wave 210 projected by the radiation source 202 for temperature change of the sample. The parameters of the electromagnetic wave 210 controlled by the controller 204 include, but are not limited to, presence, intensity, wavelength, angle of incidence, and any combination thereof. In some embodiments, the controller is manually operated, for example, it simply controls the on and off of the radiation source like a manual switch, thereby controlling the presence of the electromagnetic wave projected by the radiation source. In other embodiments, the controller includes hardware and software configured to automatically control the electromagnetic wave according to one or more predetermined programs.

[0085] In some embodiments, the predetermined program refers to a schedule in which the parameters (e.g., presence, intensity, and / or wavelength) of the electromagnetic wave 210 are set to predetermined levels for corresponding predetermined time periods. In other embodiments, the predetermined program refers to a schedule in which the temperature of the sample 90 is set to predetermined levels for corresponding predetermined time periods, and the time periods for the sample temperature to change from one predetermined level to another predetermined level are also set separately. In some embodiments, the controller 204 is configured to be programmable, which means that the controller 204 includes hardware and software configured to receive and execute a predetermined program of the system, which is communicated by an operator of the system.

[0086] Thermal cycler systems and related methods of the present invention are used to promote chemical, biological or medical determinations or reactions. In some embodiments, the reaction requires a temperature change. In some embodiments, the reaction requires or preferably a rapid temperature change to avoid non-specific reactions and / or reduce waiting time. In certain embodiments, the systems and methods of the present invention are used to promote reactions that require cyclic temperature changes to amplify nucleotides in a fluid sample; these reactions include but are not limited to polymerase chain reactions (PCR). The following description uses PCR as an example to illustrate the performance and use of the thermal cycler systems and methods of the present invention. However, it should be noted that some embodiments of the devices, systems and methods described herein are also applicable to other determinations and / or reactions that require temperature control and change.

[0087] refer to Figure 2 In some embodiments, the sample 90 is a premixed reaction medium for polymerase chain reaction (PCR). For example, in some embodiments, the reaction medium includes components such as, but not limited to: a DNA template, two primers, a DNA polymerase (e.g., Taq polymerase), deoxynucleoside triphosphates (dNTPs), divalent cations (e.g., Mg 2+ ), monovalent cations (such as K + The specific components, concentrations of the components, and total volume vary according to the rational design of the reaction.

[0088] In some embodiments, a PCR assay requires a number of changes / alterations in sample temperature between the following steps: (i) an optional initialization step, which requires heating the sample to 92-98° C.; (2) a denaturation step, which requires heating the sample to 92-98° C.; (3) an annealing step, which requires lowering the sample temperature to 50-65° C.; (4) an extension (or elongation) step, which requires heating the sample to 75-80° C.; (5) repeating steps (2)-(4) about 20-40 times; and (6) completing the assay and lowering the sample temperature to ambient temperature (e.g., room temperature) or cooling to about 4° C. The specific temperature and specific time period for each step vary and depend on many factors, including but not limited to the length of the target sequence, the length of the primers, the cation concentration, and / or the GC percentage.

[0089] The thermal cycler system of the present invention enables rapid temperature changes in PCR assays. See, for example, Figure 1 Figures (A) and (B) and Figure 2(B), in some embodiments, a sample 90 (e.g., a premixed reaction medium) is added to one or both of the plates 10 and 20 in an open configuration, and the plates are switched to a closed configuration to press the sample 90 into a thin layer having a thickness 102, wherein the thickness 102 is adjusted by a spacing mechanism (not shown); a radiation source 202 projects an electromagnetic wave 210 onto the first plate 10 (e.g., specifically onto the radiation absorbing layer 112); the radiation absorbing layer 112 is configured to absorb the electromagnetic wave 210 and convert at least a majority of the electromagnetic wave 210 into heat, which increases the temperature of the sample; removal of the electromagnetic wave 210 causes the temperature in the sample 90 to decrease.

[0090] In some embodiments, by projecting electromagnetic waves 210 onto the radiation absorbing layer 112 or increasing the intensity of the electromagnetic waves, the thermal cycler system provides rapid heating (increasing the temperature) for any or all of the initialization step, the denaturation step, and / or the extension / extension step; in some embodiments, by removing the electromagnetic waves projected by the radiation source 202 or reducing the intensity of the electromagnetic waves, the cooling in the annealing step and / or the final cooling step is quickly achieved. In some embodiments, the electromagnetic wave 210 or the increase in the intensity of the electromagnetic wave 210 causes the temperature rate of the rise to be at least 50°C / s, 45°C / s, 40°C / s, 35°C / s, 30°C / s, 25°C / s, 20°C / s, 18°C / s, 16°C / s, 14°C / s, 12°C / s, 10°C / s, 9°C / s, 8°C / s, 7°C / s, 6°C / s, 5°C / s, 4°C / s, 3°C / s, or 2°C / s, or in a range between any one of the two values. In certain embodiments, the average heating rate in the PCR assay is 10°C / s or more. In certain embodiments, the removal of electromagnetic wave 210 or the reduction in the intensity of electromagnetic wave 210 causes the temperature rate of the decline to be at least 50°C / s, 45°C / s, 40°C / s, 35°C / s, 30°C / s, 25°C / s, 20°C / s, 18°C / s, 16°C / s, 14°C / s, 12°C / s, 10°C / s, 9°C / s, 8°C / s, 7°C / s, 6°C / s, 5°C / s, 4°C / s, 3°C / s, or 2°C / s, or in the range between any one of the two values. In certain embodiments, the average cooling rate in the PCR assay is 5°C / s or more. As used herein, the term "rate" refers to the temperature change speed between two preset temperatures. In certain embodiments, the average rising or falling temperature of each step is different.

[0091] In the PCR process, after the target temperature has been reached in any step, the sample needs to be kept at the target temperature for a certain period of time. The thermal cycler system of the present invention provides a temperature maintenance function by: (1) adjusting the intensity of the electromagnetic wave 210, reducing the intensity of the electromagnetic wave 210 if the temperature has risen to the target, or increasing the intensity of the electromagnetic wave 210 if the temperature has fallen to the target, and / or (2) maintaining the target temperature by balancing the heat provided to the sample and the heat removed from the sample.

[0092] Figure 3 A cross-sectional view of an embodiment of the present invention is shown, illustrating a thermal cycler system and showing additional elements that facilitate temperature changes and control. Figure 2 As shown, the thermal cycler system includes a sample unit 100 and a thermal control unit 200. The sample unit 100 includes a first plate 10, a second plate 20, a spacer 40 and a seal 30; the thermal control unit 200 includes a radiation source 202, a controller 204, a thermometer 206 and an extender 208.

[0093] Figure 3 A sample unit 100 is shown in a closed configuration, wherein the inner surfaces 11 and 21 of the first and second plates 10 and 20 face each other, and the spacing 102 between the two plates is adjusted by a spacing mechanism 40. If the sample 90 has been deposited on one or both of the plates in the open configuration, when switching to the closed configuration, the first plate 10 and the second plate 20 are pressed by hand or other mechanism, and the sample 90 is thus compressed into a thin layer by the two plates. In some embodiments, the thickness of the layer is uniform and the same as the spacing 102 between the two plates. In certain embodiments, the spacing 102 (and therefore the thickness of the sample layer) is adjusted by the spacing mechanism 40. In some embodiments, the spacing mechanism includes a closed spacer fixed to one of the plates. In some embodiments, the spacing mechanism 40 includes a plurality of columnar spacers fixed to one or both of the plates. Here, the term "fixed" means that the spacer is attached to the plate and remains attached at least during the use of the plate.

[0094] In some embodiments, the sample unit 10 is a compression regulated open flow (CROF, also known as QMAX) device, such as, but not limited to, U.S. Provisional Patent Application No. 62 / 202,989 filed on August 10, 2015, U.S. Provisional Patent Application No. 62 / 218,455 filed on September 14, 2015, U.S. Provisional Patent Application No. 62 / 293,188 filed on February 9, 2016, U.S. Provisional Patent Application No. 62 / 305,123 filed on March 8, 2016, U.S. Provisional Patent Application No. 62 / 369,181 filed on July 31, 2016, U.S. Provisional Patent Application No. 62 / 369,181 filed on September 15, 2016 The CROF device described in U.S. Provisional Patent Application No. 62 / 394,753 filed on August 14, 2016, PCT Application (designated U.S.) No. PCT / US2016 / 045437 filed on August 10, 2016, PCT Application (designated U.S.) No. PCT / US2016 / 051775 filed on September 14, 2016, PCT Application (designated U.S.) No. PCT / US2016 / 051794 filed on September 15, 2016, and PCT / US2016 / 054025 filed on September 27, 2016, the entire disclosures of which are incorporated herein by reference for all purposes.

[0095] In some embodiments, the sample unit 100 includes a sealing element 30 configured to seal the gap 102 between the first plate 10 and the second plate 20 outside the media contact area in a closed configuration. In some embodiments, the sealing element 30 encloses the sample 90 within a certain area (e.g., a sample receiving area) so that the entire lateral area of ​​the sample 90 is clearly defined and measurable. In some embodiments, the sealing element 30 improves the uniformity of the sample 90, especially the thickness of the sample layer.

[0096] In some embodiments, the sealing element 30 includes an adhesive applied between the first plate 10 and the second plate 20 in a closed configuration. The adhesive is selected from the following materials, such as but not limited to: starch, dextrin, gelatin, asphalt, bitumin, polyisoprene natural rubber, resin, shellac, cellulose and its derivatives, vinyl derivatives, acrylic derivatives, reactive acrylic bases, polychloroprene, styrene-butadiene, styrene-diene-styrene, polyisobutylene, acrylonitrile-butadiene, polyurethane, polysulfide, silicone, aldehyde condensation resin, epoxy resin, amino resin, polyester resin, polyolefin polymer, soluble silicate, phosphate cement, or any other adhesive material, or any combination thereof. In some embodiments, the adhesive is a drying adhesive, a pressure-sensitive adhesive, a contact adhesive, a thermal adhesive, or a single-component or multi-component reactive adhesive, or any combination thereof. In some embodiments, the adhesive is a natural adhesive or a synthetic adhesive, or from any other source, or any combination thereof. In some embodiments, the adhesive is self-curing, heat-curing, UV-curing, or cured by any other treatment, or any combination thereof.

[0097] In some embodiments, the sealing element 30 includes a closed spacer (hole). For example, from a top view, the closed spacer has a circular shape (or any other closed shape) and surrounds the sample 90, substantially defining the sample 90 with the first plate 10 and the second plate 20. In certain embodiments, the closed spacer (hole) also serves as the spacing mechanism 40. In such an embodiment, the closed spacer seals the lateral boundaries of the sample 90 and adjusts the thickness of the sample layer.

[0098] In some embodiments, the controller 204 is configured to adjust the temperature of the sample according to a predetermined program to facilitate an assay and / or reaction involving the sample 90. In some embodiments, the assay and / or reaction is PCR. In certain embodiments, the controller 204 is configured to control the presence, intensity, and / or frequency of electromagnetic waves from the radiation source 206.

[0099] like Figure 3 As shown, in some embodiments, the thermal control unit 200 includes a thermometer 206. In some embodiments, the thermometer 206 provides a monitoring and / or feedback mechanism to control / monitor / regulate the temperature of the sample 90. For example, in some embodiments, the thermometer 206 is configured to measure the temperature at or near the sample contact area. In certain embodiments, the thermometer 206 is configured to directly measure the temperature of the sample 90. In some embodiments, the thermometer 206 is selected from the group consisting of: an optical fiber thermometer, an infrared thermometer, a liquid crystal thermometer, a pyrometer, a quartz thermometer, a silicon bandgap temperature sensor, a temperature bar, a thermistor, and a thermocouple. In certain embodiments, the thermometer 206 is an infrared thermometer.

[0100] In some embodiments, the thermometer 206 is configured to send a signal to the controller 204. Such a signal contains information related to the temperature of the sample 90, so that the controller 204 makes corresponding changes. For example, in the PCR process, for the denaturation step, the target temperature is set to 95°C; after measurement, the thermometer sends a signal to the controller 204 indicating that the measured temperature of the sample 90 is actually 94.8°C; the controller 204 therefore changes the output of the radiation source 202, which projects electromagnetic waves or adjusts specific parameters (such as intensity or frequency) of existing electromagnetic waves, so that the temperature of the sample 90 increases to 95°C. This measurement signal adjustment cycle is applied to any step in any reaction / assay.

[0101] like Figure 3 As shown, the thermal control unit 200 includes a beam expander 208, which is configured to expand the electromagnetic wave from the radiation source 202 from a smaller diameter to a larger diameter. In some embodiments, the electromagnetic wave projected from the radiation source 202 is sufficient to cover the entire sample contact area; however, in some embodiments, it is necessary to expand the coverage area of ​​the electromagnetic wave directly projected from the radiation source 202 to produce an expanded electromagnetic wave 210 to provide a heat source for all sample contact areas. The beam expander 208 adopts any known technology, including but not limited to the beam expanders described in U.S. Patent Nos. 4,545,677, 4,214,813, 4,127,828, and 4,016,504, and U.S. Patent Publication Nos. 2008 / 0297912 and 2010 / 0214659, the entire contents of which are incorporated herein by reference for all purposes.

[0102] Multiplexing

[0103] Figure 4 A perspective view of another embodiment of the present invention is shown, wherein there are multiple sample contact areas on the plate, allowing multiple samples to be processed and analyzed. Figure 3 As shown in Figures (A) and (B), the thermal cycler system of the present invention includes a sample unit 100 and a thermal control unit 200; the sample unit 100 includes a first plate 10, multiple second plates 20 and multiple spacing mechanisms (not shown); the thermal control unit 200 includes a radiation source 202 and a controller 204.

[0104] refer to Figure 4 In FIG. 1A , one or both of the plates (eg, the first plate 10 ) include multiple sample contact areas (not labeled). In some embodiments, one or both of the plates (eg, the first plate 10 ) include multiple radiation absorbing layers 112 . Figure 4Figure (A) shows a sample unit 100 in an open configuration, wherein the first plate 10 and the second plate 20 are partially or completely separated, allowing one or more samples to be deposited on one or both of the plates. In the open configuration, the spacing between the first plate 10 and the second plate 20 is not adjusted by a spacing mechanism.

[0105] Figure 4 Figure (B) shows a sample unit 100 in a closed configuration, wherein the inner surfaces of the two plates face each other and the spacing 102 between the two plates is adjusted by a spacing mechanism (not shown). If one or more samples have been deposited on the plates, the plates are configured to press each sample into a layer, the thickness of the layer being adjusted by the spacing mechanism.

[0106] like Figure 4 As shown in FIG. (B), a plurality of second plates 20 are used to cover a portion of the first plate 10. For example, each second plate 20 covers a single sample contact area on which a sample is deposited. There is a spacing mechanism for each sample contact area, and the spacing mechanism has a different height, thereby generating a different spacing 102 for each sample contact area and for different thicknesses of each sample layer. For example, the spacing mechanism is a columnar spacer; each sample contact area has a set of spacers with uniform height; the heights of different sets of spacers are the same or different, resulting in the same or different sample layer thicknesses for different samples.

[0107] See also Figure 4 In some embodiments, the controller 204 directs the radiation source 202 to project the electromagnetic wave 210 onto the first plate 10 (and thus onto the radiation absorbing layer 112), wherein the electromagnetic wave 210 is absorbed by the radiation absorbing layer 112 and converted into heat, resulting in a temperature change in the sample. In some embodiments, when there are multiple sample contact areas, multiple samples are processed and analyzed. For example, in some embodiments, each sample is a premixed PCR reaction medium with different components. A sample unit 100 is used to test different conditions for amplifying the same nucleotide and / or amplifying different nucleotides under the same or different conditions.

[0108] Exemplary Embodiments

[0109] Figure 5 A cross-sectional view of an exemplary process for nucleic acid amplification using a QMAX card device is shown. An example of the steps includes (A) introducing a sample containing nucleic acid to the inner side of a first plate (base plate); (B) pressing a second plate (QMAX card) onto the inner surface of the first plate to form a closed configuration of the device, wherein necessary reagents for nucleic acid amplification are dried on the inner surface of the second plate; (c) accumulating nucleic acid amplification products in a chamber enclosed by the first and second plates.

[0110] Figure 5A cross-sectional view of an exemplary process for nucleic acid amplification using a QMAX card device is shown.

[0111] More specifically, in step (A), "sample" can be any nucleic acid containing or not containing sample, including but not limited to human body fluids, such as whole blood, plasma, serum, urine, saliva and sweat, and cell culture (mammals, plants, bacteria, fungi). The sample can be freshly obtained, or stored or processed in any desired or convenient manner, such as by dilution or addition of buffers, or other solutions or solvents. Cell structures, such as human cells, animal cells, plant cells, bacterial cells, fungal cells, and virus particles, can be present in the sample.

[0112] The term "nucleic acid" as used herein refers to any DNA or RNA molecule, or a DNA / RNA hybrid, or a mixture of DNA and / or RNA. Therefore, the term "nucleic acid" includes but is not limited to genomic or chromosomal DNA, plasmid DNA, amplified DNA, cDNA, total RNA, mRNA and small RNA. The term "nucleic acid" is also intended to include natural DNA and / or RNA molecules, or synthetic DNA and / or RNA molecules. In certain embodiments, there is cell-free nucleic acid in the sample, as used herein, "cell-free" means that nucleic acid is not included in any cell structure. In some other embodiments, nucleic acid is included in the cell structure, which includes but is not limited to human cells, animal cells, plant cells, bacterial cells, fungal cells, and / or viral particles. There may be nucleic acid in the form of cell-free nucleic acid or in the cell structure or in its combined form in the sample. In some other embodiments, nucleic acid is purified before being introduced on the inner surface of the first plate. In further embodiments, nucleic acid can be in a complex combined with other molecules (such as proteins and lipids).

[0113] The method of the present invention is applicable to samples with a certain volume range. Samples with different volumes can be introduced into plates with different sizes.

[0114] If desired, the sample can be introduced onto the first plate or the second plate, or even both. Figure 5 Embodiments are provided in which the sample is introduced onto the inner surface of the first plate.

[0115] More specifically, in step (b), the second plate is pressed onto the inner surface of the first plate, in contact with the sample, to form a closed configuration of the device. As used herein, "second plate" refers to a QMAX card having periodic spacers on the inner surface that contacts the sample.

[0116] As used herein, "nucleic acid amplification" includes any technique for detecting nucleic acids by amplifying (producing a large number of copies of) a target molecule in a sample, where "target" refers to a sequence or partial sequence of a nucleic acid of interest. Suitable nucleic acid amplification techniques include, but are not limited to, different polymerase chain reaction (PCR) methods, such as hot start PCR, nested PCR, gradient PCR, reverse transcription PCR, RACEPCR, digital PCR, etc., and isothermal amplification methods, such as loop-mediated isothermal amplification (LAMP), strand displacement amplification, helicase-dependent amplification, nickase amplification, rolling circle amplification, recombinase polymerase amplification, etc.

[0117] As used herein, "essential reagents" include, but are not limited to, primers, deoxynucleotides (dNTPs), divalent cations (e.g., Mg 2+ ), monovalent cations (such as K + ), buffers, enzymes, and reporter molecules. The necessary reagents for nucleic acid amplification can be in dry form on the inner surface of the first plate or the second plate or both, or in liquid form encapsulated, embedded or surrounded in a material that melts with increasing temperature (such as, for example, paraffin wax).

[0118] As used herein, "primer", in some embodiments, may refer to a pair of forward primers and reverse primers. In some embodiments, primers may refer to multiple primers or primer sets. As used herein, enzymes suitable for nucleic acid amplification include, but are not limited to, DNA-dependent polymerases, or RNA-dependent DNA polymerases, or DNA-dependent RNA polymerases.

[0119] As used herein, the term "reporter molecule" refers to any label, marker or dye that can bind to or intercalate into a nucleic acid molecule or be activated by a byproduct of an amplification process to visualize the nucleic acid molecule or the amplification process. Suitable reporter molecules include, but are not limited to, fluorescent markers or tags or dyes, intercalators, molecular beacon markers, or bioluminescent molecules, or combinations thereof.

[0120] In some other embodiments, as used herein, "essential reagents" may also include cell lysis reagents that promote the destruction of cell structures. Cell lysis reagents include, but are not limited to, salts, detergents, enzymes, and other additives. The term "salt" herein includes, but is not limited to, lithium salts (e.g., lithium chloride), sodium salts (e.g., sodium chloride), potassium salts (e.g., potassium chloride). The term "detergent" herein may be ionic, including anionic and cationic, nonionic or zwitterionic. The term "ionic detergent" used herein includes any detergent that is partially or completely in ionic form when dissolved in water. Suitable anionic detergents include, but are not limited to, sodium dodecyl sulfate (SDS) or other alkali metal alkyl sulfates or similar detergents, sarkosyl, or a combination thereof. The term "enzyme" herein includes, but is not limited to, lysozyme, cellulase, and protease. In addition, chelating agents may also be included in cell lysis reagents, including, but not limited to, EDTA, EGTA, and other polyaminocarboxylic acids, and some reducing agents, such as dithiothreitol (dTT). The composition of the essential reagents herein varies according to the rational design of different amplification reactions.

[0121] More specifically, in step (c), when the device is in a closed configuration, the radiation source projects electromagnetic waves to the radiation absorbing layer on the inner or outer surface of the first plate or the second plate or both. The radiation absorbing layer is configured to absorb electromagnetic waves and convert at least most of the energy from the electromagnetic waves into the form of heat, which is transferred to the sample in the closed chamber. In some embodiments, the radiation source is programmed to adjust the temperature of the sample within the range of ambient temperature to 98°C. In some embodiments, for example, for conventional PCR, the sample is first heated to 98°C, and then undergoes repeated cycles of 94°C, 50-65°C and 72°C for 15-40 times. In some embodiments, for example, for isothermal amplification, the temperature of the sample is maintained at a constant temperature. In some embodiments, for example, when isothermal amplification is performed by LAMP, the sample is heated to 60-65°C for about 1-70 minutes.

[0122] As used herein, "nucleic acid amplification product" refers to various nucleic acids produced by nucleic acid amplification technology. The types of nucleic acid amplification products herein include, but are not limited to, single-stranded DNA, single-stranded RNA, double-stranded DNA, linear DNA, or circular DNA, etc. In some embodiments, the nucleic acid amplification products can be the same nucleic acids with the same length and structure. In some other embodiments, the nucleic acid amplification products can be a variety of nucleic acids with different lengths and structures.

[0123] In some embodiments, a reporter molecule is used to quantify the nucleic acid accumulated after nucleic acid amplification. As defined and used above, a reporter molecule has a quantifiable characteristic that is related to the presence or absence, or amount, of the nucleic acid amplicon accumulated in the closed chamber.

[0124] Figure 6 A cross-sectional view of an exemplary assay process combining nucleic acid extraction and amplification using a QMAX card device is shown. Examples of steps include (a) immobilizing capture probes on the inner surface of a first plate (substrate); (b) introducing a sample onto the inner surface of the first plate; (c) pressing a second plate (QMAX card 1) onto the inner surface of the first plate to form a closed configuration of the device, wherein a necessary reagent 1 that facilitates release and capture of nucleic acids is dried on the inner surface of the second plate; (d) capturing nucleic acids from the above sample onto the inner surface of the first plate; (e) removing the second plate and cleaning the inner surface of the first plate with a sponge; (f) pressing a third plate (QMAX card 2) onto the inner surface of the first plate, wherein a necessary reagent 2 for nucleic acid amplification is dried on the inner surface of the third plate; (g) accumulating nucleic acid amplification products in a chamber enclosed by the first and third plates.

[0125] More specifically, in step (a), the capture probe is fixed on the inner surface of the first plate. As used herein, "capture probe" refers to an oligonucleotide with a length of 1-200bp, preferably 5-50bp, and more preferably 10-20bp. The capture probe has a sequence complementary to the nucleic acid sequence of interest in the sample. In some embodiments, the same capture probe is fixed on the surface of the first plate. In some other embodiments, different capture probes with different base pair compositions are fixed on the surface of the first plate. The capture probe can be DNA, or RNA, or both, but preferably single-stranded DNA. As used herein, "fixation" refers to the process of anchoring the capture probe on the plate surface. In some embodiments, the capture probe is anchored by a covalent bond, wherein, for example, the 5' or 3' end of the capture probe is modified to promote coating on the plate surface. Commonly used 3' end modifications include but are not limited to thiols, dithiols, amines, biotin, etc. In some other embodiments, the capture probe can be passively absorbed on the substrate surface.

[0126] After immobilization with the capture probe, the plate surface is blocked with a blocking agent solution. Suitable blocking agents include, but are not limited to, 6-mercapto-hexanol, bovine serum albumin, and the like.

[0127] like Figure 6 As shown in step (b), "sample" can be any nucleic acid containing or not containing the sample, including but not limited to human body fluids, such as whole blood, plasma, serum, urine, saliva and sweat, and cell culture (mammals, plants, bacteria, fungi). The sample can be freshly obtained, or stored or processed in any desired or convenient manner, such as by dilution or addition of a buffer, or other solution or solvent. Cell structures such as human cells, animal cells, plant cells, bacterial cells, fungal cells, and viral particles can be present in the sample.

[0128] The term "nucleic acid" as used herein refers to any DNA or RNA molecule, or a DNA / RNA hybrid, or a mixture of DNA and / or RNA. Therefore, the term "nucleic acid" includes but is not limited to genomic or chromosomal DNA, plasmid DNA, amplified DNA, cDNA, total RNA, mRNA and small RNA. The term "nucleic acid" is also intended to include natural DNA and / or RNA molecules, or synthetic DNA and / or RNA molecules. In certain embodiments, there is cell-free nucleic acid in the sample, as used herein, "cell-free" means that nucleic acid is not included in any cell structure. In some other embodiments, nucleic acid is included in the cell structure, which includes but is not limited to human cells, animal cells, plant cells, bacterial cells, fungal cells, and / or viral particles. There may be nucleic acid in the form of cell-free nucleic acid or in the cell structure or in its combined form in the sample. In some other embodiments, nucleic acid is purified before being introduced on the inner surface of the first plate. In further embodiments, nucleic acid can be in a complex combined with other molecules (such as proteins and lipids).

[0129] The method of the present invention is applicable to samples with a certain volume range. Samples with different volumes can be introduced into plates with different sizes.

[0130] If desired, the sample can be introduced onto the first plate or the second plate, or even both. Figure 6 An example is provided in which the sample is introduced onto the inner surface of the first plate.

[0131] More specifically, in step (c), the second plate (QMAX card 1) is pressed onto the inner surface of the first plate (base plate) in contact with the sample to form a closed configuration of the device. The necessary reagent 1 for nucleic acid amplification is in dry form on the inner surface of the first plate or the second plate or both, or in liquid form encapsulated, embedded or surrounded in a material that melts with increasing temperature (e.g., paraffin).

[0132] As used herein, "essential reagent 1" refers to a cell lysis reagent, or a hybridization reagent, or a combination thereof.

[0133] As used herein, "cell lysis reagents" include, but are not limited to, salts, detergents, enzymes, and other additives that promote the destruction of cell structures. The term "salts" herein includes, but is not limited to, lithium salts (e.g., lithium chloride), sodium salts (e.g., sodium chloride), potassium salts (e.g., potassium chloride). The term "detergent" herein may be ionic, including anionic and cationic, nonionic or zwitterionic. The term "ionic detergent" used herein includes any detergent that is partially or completely in ionic form when dissolved in water. Suitable anionic detergents include, but are not limited to, sodium dodecyl sulfate (SDS) or other alkali metal alkyl sulfates or similar detergents, sarkosyl, or a combination thereof. The term "enzyme" herein includes, but is not limited to, lysozyme, cellulase, and protease. In addition, chelating agents may also be included in cell lysis reagents, including, but not limited to, EDTA, EGTA, and other polyaminocarboxylic acids, and some reducing agents, such as dithiothreitol (dTT). The composition of the necessary reagents herein varies according to the rational design of different amplification reactions.

[0134] As used herein, "hybridization reagent" refers to a reagent that promotes hybridization between the immobilized capture probe and the target nucleic acid in the sample, including but not limited to sodium chloride, sodium acetate, polysucrose, dextran, polyvinyl pyrrolidone, bovine serum albumin, etc.

[0135] More specifically, in step (d), after contacting with the above-mentioned sample, the dried necessary reagent 1 is dissolved in the sample. The target nucleic acid released from the destroyed cell structure or present as cell-free nucleic acid or a combination thereof is hybridized with the complementary capture probe on the plate surface. The time used for hybridization varies greatly depending on the specifications of the spacer on the inner surface of the QMAX card 1. In some embodiments, for example, when a QMAX card 1 with a spacer having a height of 30 μm is used, the experimental data indicates that the hybridization between the target nucleic acid and the fixed capture probe reaches equilibrium after 2 minutes. As used herein, Figure 6 In (d), "unhybridized nucleic acid" refers to nucleic acid that is not captured by the immobilized capture probe.

[0136] More specifically, in step (e), the second plate (QMAX card 1) is separated from the first plate (substrate), and the surface of the first plate (substrate) is cleaned using a sponge. As used herein, "sponge" refers to a class of flexible porous materials that change pore size under different pressures. A sponge containing a wash buffer is contacted with the surface of the first plate to remove contaminants. In some embodiments, the sponge contacts the surface of the first plate once. In some other embodiments, the sponge contacts the surface of the first plate twice or more. As used herein, "contaminants" refer to compounds that are not conducive to nucleic acid amplification reactions, including but not limited to cell debris, proteins, non-specific nucleic acids, etc.

[0137] More specifically, in step (f), the third plate (QMAX card 2) is pressed onto the inner surface of the first plate, in contact with the sample, to form a closed configuration of the device. The necessary reagents 2 for nucleic acid amplification may be in dry form on the inner surface of the first plate or the third plate or both, or in liquid form encapsulated, embedded or surrounded in a material that melts with increasing temperature (such as, for example, paraffin wax).

[0138] As used herein, "nucleic acid amplification" includes any technique for detecting nucleic acids by amplifying (producing a large number of copies of) a target molecule in a sample, where "target" refers to a sequence or partial sequence of a nucleic acid of interest. Suitable nucleic acid amplification techniques include, but are not limited to, different polymerase chain reaction (PCR) methods, such as hot-start PCR, nested PCR, gradient PCR, reverse transcription PCR, RACE PCR, digital PCR, etc., and isothermal amplification methods, such as loop-mediated isothermal amplification (LAMP), strand displacement amplification, helicase-dependent amplification, nickase amplification, rolling circle amplification, recombinase polymerase amplification, etc.

[0139] As used herein, "essential reagent 2" includes, but is not limited to, primers, deoxynucleotides (dNTPs), divalent cations (e.g., Mg 2+ ), monovalent cations (such as K + ), buffer, enzyme, and reporter molecule. The necessary reagent 2 for nucleic acid amplification can be in dry form on the inner surface of the first plate or the second plate or both, or in liquid form encapsulated, embedded or surrounded in a material that melts with increasing temperature (such as, for example, paraffin).

[0140] As used herein, "primer", in some embodiments, may refer to a pair of forward primers and reverse primers. In some embodiments, primers may refer to multiple primers or primer sets. As used herein, enzymes suitable for nucleic acid amplification include, but are not limited to, DNA-dependent polymerases, or RNA-dependent DNA polymerases, or DNA-dependent RNA polymerases.

[0141] As used herein, the term "reporter molecule" refers to any label, marker or dye that can bind to or intercalate into a nucleic acid molecule or be activated by a byproduct of an amplification process to visualize the nucleic acid molecule or the amplification process. Suitable reporter molecules include, but are not limited to, fluorescent markers or tags or dyes, intercalators, molecular beacon markers, or bioluminescent molecules, or combinations thereof.

[0142] More specifically, in step (g), when the device is in a closed configuration, the radiation source projects electromagnetic waves to the radiation absorbing layer on the inner or outer surface of the first plate or the third plate or both. The radiation absorbing layer is configured to absorb electromagnetic waves and convert at least most of the energy from the electromagnetic waves into the form of heat, which is transferred to the sample in the closed chamber. In some embodiments, the radiation source is programmed to adjust the temperature of the sample within the range of ambient temperature to 98°C. In some embodiments, for example, for conventional PCR, the sample is first heated to 98°C, and then undergoes repeated cycles of 94°C, 50-65°C and 72°C for 15-40 times. In some embodiments, for example, for isothermal amplification, the temperature of the sample is maintained at a constant temperature. In some embodiments, for example, when isothermal amplification is performed by LAMP, the sample is heated to 60-65°C for about 1-70 minutes.

[0143] As used herein, "nucleic acid amplification product" refers to various nucleic acids produced by nucleic acid amplification technology. The types of nucleic acid amplification products herein include, but are not limited to, single-stranded DNA, single-stranded RNA, double-stranded DNA, linear DNA, or circular DNA, etc. In some embodiments, the nucleic acid amplification products can be the same nucleic acids with the same length and structure. In some other embodiments, the nucleic acid amplification products can be a variety of nucleic acids with different lengths and structures.

[0144] In some embodiments, a reporter molecule is used to quantify the nucleic acid accumulated after nucleic acid amplification. As defined and used above, a reporter molecule has a quantifiable characteristic that is related to the presence or absence, or amount, of the nucleic acid amplicon accumulated in the closed chamber.

[0145] In some embodiments of the QMAX, the sample contact area of ​​one or both plates contains a compressive open flow monitoring surface structure (MSS) that is configured to monitor how much flow has occurred after the COF. For example, in some embodiments, the MSS contains a shallow square array that will cause friction to components in the sample (e.g., blood cells in blood). By examining the distribution of some components of the sample, information related to the flow of the sample and its components under the COF can be obtained.

[0146] The depth of the MSS may be 1 / 1000, 1 / 100, 1 / 100, 1 / 5, 1 / 2 of the spacer height, or within the range of any two values, and be in the shape of a protrusion or a hole.

[0147] In some embodiments, a device for measuring a thin fluid sample layer comprises:

[0148] A first plate, a second plate, a spacer and a fixture, wherein:

[0149] i. The first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0150] ii. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample;

[0151] iii. One or both of the plates contain spacers fixed to the respective plates;

[0152] iv. the spacers have a predetermined substantially uniform height equal to or less than 200 microns, wherein at least one of the spacers is within the sample contact area; and

[0153] v. The clamp compresses the first plate and the second plate to fix the two plates together in a closed configuration, wherein the pressure of the clamp inserted on the plate is 0.01 kg / cm^2 or more,

[0154] wherein in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates; and

[0155] wherein in a closed configuration configured after the sample is deposited in an open configuration, at least a portion of the sample is compressed by two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and regulated by the plates and spacers.

[0156] In some embodiments, a device for rapidly changing the temperature of a thin fluid sample layer comprises:

[0157] A first plate, a second plate, a spacer and a fixture, wherein:

[0158] vi. The first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0159] vii. each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample,

[0160] viii. One or both of the plates include a spacer fixed to the respective plate;

[0161] ix. The spacers have a predetermined substantially uniform height equal to or less than 200 microns, wherein at least one of the spacers is within the sample contact area; and

[0162] x. A clamp compresses the first plate and the second plate to fix the two plates together in a closed configuration, wherein the pressure of the clamp inserted on the plate is 0.01 kg / cm^2 or more,

[0163] wherein in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates; and

[0164] wherein in a closed configuration configured after the sample is deposited in the open configuration, at least a portion of the sample is compressed by the two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and regulated by the plates and the spacers; and

[0165] The plates and fixtures are configured to allow the temperature of the sample to change at a rate of 10°C / s or greater.

[0166] In some embodiments, a device for rapidly changing the temperature of a thin fluid sample layer comprises: a first plate, a second plate, and a spacer, wherein:

[0167] iii. The first plate has a thickness of 100 μm (micrometers) or less;

[0168] iv. the first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0169] v. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample; and

[0170] vi. One or both of the plates contain spacers fixed to the respective plates;

[0171] vii. The spacers have a predetermined substantially uniform height equal to or less than 200 microns;

[0172] viii. At least one of the spacers is located in the sample contact area;

[0173] wherein in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates; and

[0174] wherein in a closed configuration configured after the sample is deposited in the open configuration, at least a portion of the sample is compressed by the two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and regulated by the plates and the spacers; and

[0175] Wherein the plate and the fixture are configured to allow the temperature of at least a portion of the sample to change at a rate of 10° C. / s or greater.

[0176] In some embodiments, a device for rapidly changing the temperature of a thin fluid sample layer comprises:

[0177] A first plate, a second plate and a fixture, wherein:

[0178] i. The first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0179] ii. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample to be assayed; and

[0180] iii. The clamp compresses the first plate and the second plate to fix the plates in a closed configuration, wherein the pressure of the clamp inserted on the plate is 0.01 kg / cm^2 or more,

[0181] wherein in the open configuration the two plates are partially or completely separated, the average spacing between the plates is 250 μm or more, and the sample is deposited on one or both of the plates; and

[0182] wherein in a closed configuration configured after the sample is deposited in an open configuration, at least a portion of the sample is compressed by two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and is 200 μm thick or less.

[0183] In some embodiments, the device of any of the preceding embodiments further comprises a radiation absorbing layer, the radiation absorbing layer being located adjacent to at least a portion of the sample of uniform thickness, and the area of ​​at least a portion of the sample and the radiation absorbing layer being substantially greater than the uniform thickness.

[0184] In some embodiments, the device of any preceding embodiment, wherein at least a portion of the sample and an area of ​​the radiation absorbing layer are substantially greater than a uniform thickness of the sample.

[0185] In some embodiments, the device has one of the plates having a thickness of 100 μm or less.

[0186] In some embodiments, further comprising a radiation absorbing layer adjacent at least a portion of the uniform thickness sample, wherein the device has one of the plates having a thickness of 100 μm or less.

[0187] In some embodiments, further comprising a clamp compressing the first plate and the second plate together in a closed configuration, wherein the pressure of the clamp inserted on the plates is 0.01 kg / cm^2 or higher.

[0188] In some embodiments, a fixture is further included to compress the first plate and the second plate together in a closed structure, and further includes a radiation absorbing layer near at least a portion of the sample of uniform thickness, wherein the pressure of the fixture inserted on the plates is 0.01 kg / cm^2 or higher.

[0189] In some embodiments, a system for rapidly changing the temperature of a thin layer of a fluid sample comprises:

[0190] i. A device as in any of the preceding embodiments,

[0191] ii. a radiation source, wherein the radiation source is configured to radiate electromagnetic waves that are significantly absorbed by the radiation absorbing layer; and

[0192] iii. The controller is configured to control the radiation source and vary the temperature of the sample.

[0193] In some embodiments, a method for rapidly changing the temperature of a thin fluid sample layer comprises:

[0194] i. Provide a device or system as in any of the preceding embodiments;

[0195] ii. depositing a fluid sample on one or both of the plates of the device;

[0196] iii. after ii, pressing the plate into a closed configuration, wherein the plate compresses at least a portion of the sample into a thin layer having a thickness of less than 200 μm; and

[0197] iv. Changing and maintaining the temperature of the sample layer by varying the presence, intensity, wavelength, frequency and / or angle of electromagnetic waves from the radiation source.

[0198] In some embodiments, the device, system, or method of any preceding embodiment, wherein the fixture is configured to include a thermal insulator layer to reduce heat conduction between the fixture and the plate, wherein the thermal insulator layer includes a material with a thermal conductivity of 2 W / mK.

[0199] In some embodiments, a device, system, or method as described in any of the preceding embodiments, wherein the fixture is configured to include a thermal insulator layer to reduce the thermal mass required to heat or cool the sample, wherein the thermal insulator layer includes a material with a thermal conductivity of 2 W / mK.

[0200] In some embodiments, the device, system, or method of any preceding embodiment, wherein in the closed configuration, the clamp is configured to seal all QMAX cards.

[0201] In some embodiments, the device, system, or method of any preceding embodiment, wherein in a closed configuration, the clamp is configured to have thermally conductive contact with a portion of the surface of the plate.

[0202] In some embodiments, the device, system or method of any preceding embodiment, wherein in the closed configuration, the clamp has thermally conductive contact only with a peripheral surface area of ​​the plate.

[0203] In some embodiments, a device, system or method as described in any of the preceding embodiments, wherein in a closed configuration, the clamp has thermally conductive contact only with a surface area of ​​the plate, wherein the surface area is outside the portion of the sample to be amplified nucleic acid.

[0204] In some embodiments, the device, system or method as described in any of the preceding embodiments, wherein the fixture includes a transparent window that allows external light to reach the plate or allows internal light to escape.

[0205] In some embodiments, a device, system or method as described in any of the preceding embodiments, wherein the fixture comprises a transparent window that allows external light to reach the plate or allows internal light to leave, wherein the transparency is greater than 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or within a range between any two values.

[0206] In some embodiments, the device, system or method of any of the preceding embodiments, wherein the clamp inserts a pressure to compress the first plate and the second plate, wherein the pressure is 0.01 kg / cm 2 , 0.1kg / cm 2 , 0.5kg / cm 2 , 1kg / cm 2 , 2kg / cm 2 、kg / cm 2 , 5kg / cm 2 , 10kg / cm 2 , 20kg / cm 2 、30kg / cm 2 , 40kg / cm 2 , 50kg / cm 2 、60kg / cm 2 , 100kg / cm 2 、150kg / cm 2 , 200kg / cm 2 , 400kg / cm 2 , or in the range between any two values.

[0207] In some embodiments, the device, system or method of any of the preceding embodiments, wherein the clamp inserts a pressure to compress the first plate and the second plate, wherein the pressure is 0.1 kg / cm 2 Up to 20kg / cm 2 .

[0208] In some embodiments, the device, system or method of any of the preceding embodiments, wherein the clamp inserts a pressure to compress the first plate and the second plate, wherein the pressure is 0.1 kg / cm 2 Up to 20kg / cm 2 .

[0209] In some embodiments, the device, system or method of any of the preceding embodiments, wherein the clamp inserts a pressure to compress the first plate and the second plate, wherein the pressure is 0.5 kg / cm 2 Up to 40kg / cm 2 .

[0210] In some embodiments, a device, system or method as described in any of the foregoing embodiments further comprises a clamp which compresses the first plate and the second plate together in the closed structure, and further comprises a sealing material which is located between at least a portion of the first plate and the second plate, wherein the pressure of the clamp inserted into the plates is 0.01 kg / cm^2 or higher.

[0211] In some embodiments, the device, system or method of any preceding embodiment, wherein the spacer has a substantially flat top.

[0212] In some embodiments, the device, system or method of any preceding embodiment, wherein one of the plates is 50 μm or less. In some embodiments, the device, system or method of any preceding embodiment, wherein changing the temperature of the sample is a thermal cycle that changes the temperature up and down in a cyclic manner.

[0213] In some embodiments, the device, system or method of any preceding embodiment, wherein changing the temperature of the sample is thermal cycling, wherein the thermal cycling is used for nucleic acid amplification using polymerase chain reaction (PCR).

[0214] In some embodiments, the device, system or method of any preceding embodiment, wherein the temperature of the sample is changed for isothermal nucleic acid amplification.

[0215] In some embodiments, the apparatus, system or method of any preceding embodiment, at least a portion of the sample and the radiation absorbing layer have an area of ​​substantially greater than uniform thickness.

[0216] In some embodiments, the device, system or method as described in any of the preceding embodiments, wherein the radiation absorbing layer comprises a disk coupled dot-pole antenna (D2PA) array, a silicon interlayer, graphene, a superlattice or other plasmonic material, or a combination thereof.

[0217] In some embodiments, the device, system, or method as described in any of the preceding embodiments, wherein the radiation absorbing layer comprises carbon or black nanostructures or a combination thereof.

[0218] In some embodiments, the device, system or method as described in any preceding embodiment, wherein the radiation absorbing layer is configured to absorb radiation energy.

[0219] In some embodiments, the device, system or method as described in any of the preceding embodiments, wherein the radiation absorbing layer is configured to radiate energy in the form of heat after absorbing the radiation energy.

[0220] In some embodiments, as described in any of the above embodiments, the radiation absorbing layer is located below the sample layer and is in direct contact with the sample layer.

[0221] In some embodiments, the device, system or method of any preceding embodiment, wherein the radiation absorbing layer is configured to absorb electromagnetic waves selected from the group consisting of radio waves, microwaves, infrared waves, visible light, ultraviolet waves, X-rays, gamma rays, and thermal radiation.

[0222] In some embodiments, the device, system or method as described in any preceding embodiment, wherein at least one of the panels does not block radiation absorbed by the radiation absorbing layer.

[0223] In some embodiments, the device, system, or method of any preceding embodiment, wherein one or both of the plates have low thermal conductivity.

[0224] In some embodiments, the device, system or method of any preceding embodiment, wherein the uniform thickness of the sample layer is regulated by one or more spacers affixed to one or both of the plates.

[0225] In some embodiments, the device, system or method of any preceding embodiment, wherein the sample is a premixed polymerase chain reaction (PCR) medium.

[0226] In some embodiments, the device, system or method of any preceding embodiment, wherein the device is configured to facilitate a PCR assay for varying the temperature of a sample according to a predetermined program.

[0227] In some embodiments, the device, system, or method as described in any of the preceding embodiments, wherein the device is configured to perform diagnostic testing, health monitoring, environmental testing, and / or forensic testing.

[0228] In some embodiments, a device, system or method as described in any of the preceding embodiments, wherein the device is configured to perform DNA amplification, DNA quantification, selective DNA isolation, genetic analysis, tissue typing, oncogene identification, infectious disease testing, genetic fingerprint analysis, and / or paternity testing.

[0229] In some embodiments, the device of any preceding embodiment, wherein the sample layer is laterally sealed to reduce sample evaporation.

[0230] In some embodiments, a system as in any embodiment further comprises a controller configured to control the presence, intensity, wavelength, frequency, and / or angle of the electromagnetic waves.

[0231] In some embodiments, the system of any preceding embodiment further comprises a thermometer configured to measure a temperature at or near the sample contact area and to send a signal to the controller based on the measured temperature.

[0232] In some embodiments, a system or method as in any of the preceding embodiments, wherein the thermometer is selected from the group consisting of: a fiber optic thermometer, an infrared thermometer, a liquid crystal thermometer, a pyrometer, a quartz thermometer, a silicon bandgap temperature sensor, a temperature strip, a thermistor, and a thermocouple.

[0233] In some embodiments, a system or method as in any preceding embodiment, wherein the controller is configured to control the presence, intensity, wavelength, frequency, and / or angle of electromagnetic waves from the radiation source.

[0234] In some embodiments, a system or method as in any of the preceding embodiments, wherein the radiation source and the radiation absorbing layer are configured such that the electromagnetic wave causes an average heating rate of at least 10°C / s; and removal of the electromagnetic wave causes an average cooling rate of at least 5°C / s.

[0235] In some embodiments, the device, system or method of any of the preceding embodiments, wherein the radiation source and the radiation absorbing layer are configured to produce an average heating rate of at least 10° C. / s and an average cooling rate of at least 5° C. / s.

[0236] In some embodiments, a device, system or method such as any of the foregoing embodiments, wherein the radiation source and the radiation absorbing layer are configured to produce an average heating rate of at least 10°C / s to achieve the initialization step, denaturation step and / or extension / extension step in the PCR process, and an average cooling rate of at least 5°C / s to achieve the annealing step and / or final cooling step in the PCR process.

[0237] In some embodiments, the device, system or method of any preceding embodiment, wherein the PCR sample comprises: template DNA, primer DNA, cation, polymerase, and buffer.

[0238] In some embodiments, the method of any of the preceding embodiments, wherein the step of pressing the panels into a closed configuration comprises pressing the panels with an imprecise pressure.

[0239] In some embodiments, the method of any of the preceding embodiments, wherein the step of pressing the panels into a closed configuration comprises directly pressing the panels with a human hand.

[0240] In some embodiments, the method of any of the preceding embodiments, wherein the thickness of the layer having a substantially uniform thickness varies by less than 10%.

[0241] In some embodiments, the device, system or method of any of the preceding embodiments further comprises a reagent selected from the group consisting of a DNA template, a primer, a DNA polymerase, deoxyribonucleoside triphosphates (dNTPs), a divalent cation (e.g., Mg 2+ ), monovalent cations (such as K + ), and a buffer solution.

[0242] In some embodiments, a device, system or method as in any of the preceding embodiments, wherein changing the temperature of the sample is thermal cycling, wherein the thermal cycling is used for nucleic acid amplification using a polymerase chain reaction (PCR), and the polymerase chain reaction (PCR) is selected from the group consisting of: hot start PCR, nested PCR, gradient PCR, reverse transcription PCR, RACE PCR, and digital PCR.

[0243] In some embodiments, a device, system or method as in any of the preceding embodiments, wherein the temperature of the sample is changed for isothermal nucleic acid amplification, and the isothermal amplification is selected from the group consisting of: loop-mediated isothermal amplification, strand displacement amplification, helicase-dependent amplification, nickase amplification, rolling circle amplification, and recombinase polymerase amplification.

[0244] Column spacer flat top

[0245] In certain embodiments of the invention, the spacer is a post having a flat top and a foot fixed to a plate, wherein the flat top has a smoothness with small surface variations, and the variation is less than 5, 10nm, 20nm, 30nm, 50nm, 100nm, 200nm, 300nm, 400nm, 500nm, 600nm, 700nm, 800nm, 1000nm, or in a range between any two values. The preferred flat post top smoothness is a surface variation of 50nm or less.

[0246] Additionally, the surface variation is relative to the spacer height, and the ratio of the column flat top surface variation to the spacer height is less than 0.5%, 1%, 3%, 5%, 7%, 10%, 15%, 20%, 30%, 40%, or within a range between any two values. Preferred flat column top smoothness has a ratio of the column flat top surface variation to the spacer height of less than 2%, 5%, or 10%.

[0247] Column spacer side wall angle

[0248] In certain embodiments of the invention, the spacers are posts having sidewall angles. In some embodiments, the sidewall angles are less than 5 degrees (measured from the normal to the surface), 10 degrees, 20 degrees, 30 degrees, 40 degrees, 50 degrees, 70 degrees, or in a range between any two values. In preferred embodiments, the sidewall angles are less than 5 degrees, 10 degrees, or 20 degrees.

[0249] Forming a uniform thin fluid layer by pressing with imprecise pressure

[0250] In certain embodiments of the present invention, a uniform thin layer of a fluid sample is formed by using a compression with an imprecise force. The term "imprecise compression force" does not add details, and then adds a definition of the imprecise compression force. As used herein, in the context of a force (e.g., "imprecise pressure"), the term "imprecise" refers to a force that is

[0251] (a) has a magnitude that is not precisely known or accurately predicted when the force is applied; (b) has a magnitude within 0.01 kg / cm 2 (square centimeters) to 100kg / cm 2 range, (c) the magnitude varies from one application of force to the next; and (d) the uncertainty (i.e., variation) in the forces in (a) and (c) is at least 20% of the total force actually applied.

[0252] Imprecise forces can be applied by the human hand, for example, by pinching objects together between the thumb and index finger, or by pinching and rubbing objects together between the thumb and index finger.

[0253] In some embodiments, the imprecise force of hand pressure is 0.01 kg / cm 2 , 0.1kg / cm 2 , 0.5kg / cm 2 , 1kg / cm 2 , 2kg / cm 2 、kg / cm 2 , 5kg / cm 2 , 10kg / cm 2 , 20kg / cm 2 、30kg / cm 2 , 40kg / cm2 , 50kg / cm 2 、60kg / cm 2 , 100kg / cm 2 、150kg / cm 2 , 200kg / cm 2 , or the range between any two values; and 0.1kg / cm 2 Up to 0.5kg / cm 2 , 0.5kg / cm 2 Up to 1kg / cm 2 , 1kg / cm 2 Up to 5kg / cm 2 , 5kg / cm 2 Up to 10kg / cm 2 (Pressure) is a preferred range of pressure.

[0254] Spacer filling factor.

[0255] The term "spacer fill factor" or "fill factor" refers to the ratio of the spacer contact area to the total panel area, where the spacer contact area is the contact area of ​​the top surface of the spacer contacting the inner surface of the panel in the closed configuration, and the total panel area is the total area of ​​the inner surface of the panel contacted by the top of the spacer. Since there are two panels and each spacer has two contact surfaces, each contacting one panel, the fill factor is the minimum fill factor.

[0256] For example, if the spacer is a column with a flat top, almost uniform cross section, and 2 μm height with a square shape (10 μm x 10 μm), and the spacer is periodic with a period of 100 μm, the filling factor of the spacer is 1%. If, in the above example, the foot of the column spacer is a square shape of 15 μm x 15 μm, the filling factor is still 1% by definition.

[0257] A method or apparatus as claimed in any preceding claim, wherein the spacer has a cylindrical shape and a substantially uniform cross-section.

[0258] A method or apparatus as claimed in any preceding claim, wherein the spacer pitch (SD) is equal to or less than about 120 μm (micrometers).

[0259] A method or apparatus as claimed in any preceding claim, wherein the spacer pitch (SD) is equal to or less than about 100 μm (micrometers).

[0260] A method or apparatus as claimed in any preceding claim, wherein the fourth power of the spacer spacing (ISD) divided by the thickness (h) and Young's modulus (E) of the flexible sheet (ISD 4 / (hE)) is 5×106 μm 3 / GPa or below.

[0261] A method or apparatus as claimed in any preceding claim, wherein the fourth power of the spacer spacing (ISD) divided by the thickness (h) and Young's modulus (E) of the flexible sheet (ISD 4 / (hE)) is 5×10 5 μm 3 / GPa or below.

[0262] A method or device as described in any preceding claim, wherein the spacers have a columnar shape, a substantially flat top surface, a predetermined substantially uniform height and a predetermined constant spacer spacing, the spacer spacing being at least about 2 times larger than the size of the analyte, wherein the Young's modulus of the spacer multiplied by the packing factor of the spacer is equal to or greater than 2 MPa, wherein the packing factor is the ratio of the spacer contact area to the total plate area, and wherein for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1 (1).

[0263] A method or apparatus as claimed in any preceding claim, wherein the spacers have a columnar shape, a substantially flat top surface, a predetermined substantially uniform height and a predetermined constant spacer spacing, the spacer spacing being at least about 2 times greater than the size of the analyte, wherein the Young's modulus of the spacers multiplied by the packing factor of the spacers is equal to or greater than 2 MPa, wherein the packing factor is the ratio of the spacer contact area to the total plate area, and wherein for each spacer, the ratio of the lateral dimension of the spacer to its height is at least one (1), wherein the fourth power of the spacer spacing (ISD) divided by the thickness (h) of the flexible plate and the Young's modulus (E) (ISD 4 / (hE)) is 5x10 6 μm 3 / GPa or below.

[0264] A device as claimed in any preceding device claim, wherein the ratio of the spacer pitch of the spacers to the average width of the spacers is 2 or greater, and the filling factor of the spacers multiplied by the Young's modulus of the spacers is 2 MPa or greater.

[0265] An apparatus or method as claimed in any preceding claim, wherein the analyte is a protein, peptide, nucleic acid, virus, bacteria, cell, nanoparticle, molecule, synthetic compound or inorganic compound.

[0266] A method or device as described in any preceding claim, wherein the sample is a biological sample selected from amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma or serum), breast milk, cerebrospinal fluid (CSF), cerumen (ear wax), chyle, chyme, endolymph, perilymph, feces, breath, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, saliva, exhaled condensate, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus and urine.

[0267] A method or apparatus as claimed in any preceding claim, wherein the spacers have a columnar shape and the ratio of the width to the height of the columns is equal to or greater than 1.

[0268] A method as claimed in any preceding claim, wherein the sample deposited on one or both plates has an unknown volume.

[0269] A method or apparatus as claimed in any preceding claim, wherein the spacers have a columnar shape, and the columns have a substantially uniform cross-section.

[0270] A method or apparatus as claimed in any preceding claim, wherein the sample is used to detect, purify and quantify compounds or biomolecules associated with certain disease stages.

[0271] A method or apparatus as claimed in any preceding claim, wherein the sample is associated with infectious and parasitic diseases, injuries, cardiovascular diseases, cancer, psychiatric disorders, neuropsychiatric disorders, pulmonary diseases, renal diseases and other and organic diseases.

[0272] A method or apparatus as claimed in any preceding claim, wherein the sample involves detection, purification or quantification of micro-organisms.

[0273] A method or apparatus as claimed in any preceding claim, wherein the sample involves viruses, fungi and bacteria from the environment (eg water, soil or a biological sample).

[0274] A method or apparatus as claimed in any preceding claim, wherein the sample relates to the detection or quantification of compounds or biological samples that pose a risk to food safety or national security (eg, toxic waste, anthrax).

[0275] A method or apparatus as claimed in any preceding claim, wherein the sample is related to the quantification of a vital parameter in medical or physiological monitoring.

[0276] A method or apparatus as claimed in any preceding claim, wherein the sample relates to glucose, blood, oxygen levels, total blood cell count.

[0277] A method or apparatus as claimed in any preceding claim, wherein the sample is associated with the detection and quantification of specific DNA or RNA from a biological sample.

[0278] A method or apparatus as claimed in any preceding claim wherein the sample is associated with sequencing and comparison of genetic sequences of chromosomal and mitochondrial DNA for genomic analysis.

[0279] A method or apparatus as claimed in any preceding claim, wherein the sample relates to detection of a reaction product, for example during synthesis or purification of a drug.

[0280] A method or apparatus as claimed in any preceding claim, wherein the sample is cells, tissue, body fluid or faeces.

[0281] A method or apparatus as claimed in any preceding claim, wherein the sample is a sample in the fields of human, veterinary, agricultural, food, environmental and pharmaceutical testing.

[0282] A method or apparatus as claimed in any preceding claim, wherein the sample is a biological sample selected from hair, nail, ear wax, breath, connective tissue, muscle tissue, neural tissue, epithelial tissue, cartilage, cancer sample or bone.

[0283] An apparatus or method as in any preceding embodiment, wherein a spacer pitch is in a range of 5 μm to 120 μm.

[0284] An apparatus or method as in any preceding embodiment, wherein the spacer pitch is in a range of 120 μm to 200 μm.

[0285] A device as claimed in any preceding device claim, wherein the flexible sheet has a thickness in the range 20 μm to 250 μm and a Young's modulus in the range 0.1 to 5 GPa.

[0286] A device as claimed in any preceding device claim, wherein for the flexible sheet, the thickness of the flexible sheet multiplied by the Young's modulus of the flexible sheet is in the range of 60 to 750 GPa-μm.

[0287] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is at least 1 mm 2 It is uniform over the lateral area.

[0288] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is at least 3 mm 2 It is uniform over the lateral area.

[0289] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is at least 5 mm 2 It is uniform over the lateral area.

[0290] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is at least 10 mm 2 It is uniform over the lateral area.

[0291] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is at least 20 mm 2 It is uniform over the lateral area.

[0292] A device as claimed in any preceding claim, wherein the uniform thickness of the sample layer is 20 mm 2 Up to 100mm 2 The lateral area within the range is uniform.

[0293] A device as claimed in any preceding device claim, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 5% or better.

[0294] A device as claimed in any preceding device claim, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 10% or better.

[0295] A device as claimed in any preceding device claim, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 20% or better.

[0296] A device as claimed in any preceding device claim, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 30% or better.

[0297] The present invention can be used for various applications in various fields where it is necessary to determine the presence and / or quantification of one or more analytes in a sample. For example, the present invention can be used to detect atoms, molecules, proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles, etc. The sample can be a sample from a variety of fields, including but not limited to humans, veterinary medicine, agriculture, food, environment, hygiene, health, beauty, etc.

[0298] Related Documents

[0299] The present invention includes various embodiments that can be combined in various ways as long as the various components do not contradict each other. The embodiments should be considered as a single invention document: each application has other applications as references and is also cited as a whole for all purposes, rather than as discrete independent files. These embodiments include not only the disclosure in the current file, but also include the files cited, incorporated or claiming priority here.

[0300] (1) definition

[0301] Terms used to describe the devices, systems, and methods disclosed herein are defined in this application or in PCT Application (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0302] The terms "CROF card (or card)", "COF card", "QMAX card", "Q card", "CROF device", "COF device", "QMAX device", "CROF board", "COF board" and "QMAX board" are interchangeable, except that in some embodiments, the COF card does not include a spacer; and these terms refer to a device that includes a first board and a second board that are movable relative to each other into different configurations (including an open configuration and a closed configuration), and the device includes a spacer that adjusts the spacing between the boards (except for some embodiments of COF). The term "X board" refers to one of the two boards in a CROF card, wherein the spacer is fixed to the board. Further description of COF cards, CROF cards and X boards are described in provisional application serial number 62 / 456065 filed on February 7, 2017, all of which are incorporated herein in their entirety for all purposes.

[0303] (2) Q-Cards, Spacers, and Uniform Sample Thickness

[0304] Devices, systems and methods disclosed herein may include or use Q cards, spacers and uniform sample thickness embodiments for sample detection, analysis and quantification. In some embodiments, the Q card includes a spacer that helps to make at least a portion of the sample a very uniform layer. Disclosed herein or in PCT applications (designated U.S.) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, the structure, materials, functions, variations and sizes of the spacer and the uniformity of the sample layer are listed, described and summarized, and the entire contents of all these applications are incorporated herein for all purposes.

[0305] (3) Hinges, opening notches, grooved edges and slides

[0306] Device, system and method disclosed herein may include or use the Q card for sample detection, analysis and quantification.In certain embodiments, Q card comprises hinge, notch, groove and slide block, and it helps to promote the operation of Q card and the measurement of sample.Disclosed herein or in PCT application (designated U.S.) No. PCT / US2016 / 045437 and PCT / US0216 / 051775, U.S. Provisional Application No. 62 / 456065, U.S. Provisional Application No. 62 / 456287, U.S. Provisional Application No. 62 / 456504 ...

[0307] (4) Q card, slider and smartphone detection system

[0308] Devices, systems and methods disclosed herein may include or use Q cards for sample detection, analysis and quantification. In certain embodiments, Q cards are used together with the slider of the card that allows the smart phone detection system to read. Disclosed herein or in PCT applications (designated U.S.) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, the structure, materials, functions, variations, sizes and connections of Q cards, sliding parts and mobile phone detection systems are listed, described and summarized, and the entire contents of all these applications are incorporated herein for all purposes.

[0309] (5) Detection Methods

[0310] Device, system and method disclosed herein may include or be used for various types of detection methods. Detection methods are disclosed herein or listed, described and summarized in PCT application (designated U.S.) No. PCT / US2016 / 045437 and PCT / US0216 / 051775, U.S. Provisional Application No. 62 / 456065, U.S. Provisional Application No. 62 / 456287, U.S. Provisional Application No. 62 / 456504, filed on February 7, 2017, respectively, filed on August 10, 2016 and September 14, 2016, the entire contents of all these applications are incorporated herein for all purposes.

[0311] (6) Label

[0312] The devices, systems and methods disclosed herein can employ various types of labels for analyte detection. Labels are disclosed herein or listed, described and summarized in PCT Applications (designated U.S.) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, the entire contents of all of which are incorporated herein for all purposes.

[0313] (7) Analytes

[0314] The devices, systems and methods disclosed herein can be used to manipulate and detect various types of analytes (including biomarkers). Analytes are disclosed herein or listed, described and summarized in PCT Application (designated US) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0315] (8) Applications (fields and samples)

[0316] Device, system and method disclosed herein can be used for various applications (fields and samples).Disclosed herein or listed, described and summarized in PCT application (designated U.S.) No. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, the entire contents of all these applications are incorporated herein for all purposes.

[0317] (9) Cloud

[0318] The devices, systems and methods disclosed herein may employ cloud technology for data transmission, storage and / or analysis. Relevant cloud technologies are disclosed herein or listed, described and summarized in PCT Applications (designated US) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, U.S. Provisional Application No. 62 / 456287, filed on February 8, 2017, and U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0319] Other Notes

[0320] Other embodiments according to the inventive subject matter of the present disclosure are described in the following enumerated paragraphs.

[0321] It must be noted that, as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise, such as when the word "single" is used. For example, reference to an "analyte" includes a single analyte and multiple analytes, reference to a "capture agent" includes a single capture agent and multiple capture agents, reference to a "detection agent" includes a single detection agent and multiple detection agents, and reference to a "reagent" includes a single reagent and multiple reagents.

[0322] As used herein, the terms "adapted" and "configured" mean that an element, component, or other subject matter is designed and / or intended to perform a given function. Thus, the use of the terms "adapted" and "configured" should not be interpreted as meaning that a given element, component, or other subject matter is simply "capable of" performing a given function. Similarly, a subject matter stated as being configured to perform a particular function may additionally or alternatively be described as being operable to perform that function.

[0323] As used herein, the phrase "for example", the phrase "as an example" and / or simply the terms "example" and "exemplary", when used with reference to one or more components, features, details, structures, embodiments and / or methods according to the present disclosure, are intended to convey that the described components, features, details, structures, embodiments and / or methods are illustrative, non-exclusive examples of components, features, details, structures, embodiments and / or methods according to the present disclosure. Therefore, the described components, features, details, structures, embodiments and / or methods are not intended to be limiting, required or exclusive / exhaustive; and other components, features, details, structures, embodiments and / or methods, including structurally and / or functionally similar and / or equivalent components, features, details, structures, embodiments and / or methods, are also within the scope of the present disclosure.

[0324] As used herein, the phrases "at least one" and "one or more" with respect to a list of more than one entity refer to any one or more entities in the list of entities, and are not limited to at least one of each and every entity specifically listed in the list of entities. For example, "at least one of A and B" (or equivalently, "at least one of A or B", or equivalently, "at least one of A and / or B") may refer to A alone, B alone, or A and B in combination.

[0325] As used herein, the term "and / or" placed between a first entity and a second entity means one of (1) the first entity, (2) the second entity, and (3) the first entity and the second entity. Multiple entities listed using "and / or" should be interpreted in the same manner, i.e., "one or more" of the entities so conjoined. In addition to the entities specifically identified by the "and / or" clause, other entities may optionally be present, whether related or unrelated to those entities specifically identified.

[0326] When numerical ranges are mentioned herein, the present invention includes embodiments in which the endpoints are included, embodiments in which both endpoints are excluded, and embodiments in which one endpoint is included and the other endpoint is excluded. Both endpoints should be assumed to be included unless otherwise stated. Furthermore, unless otherwise stated or apparent to one of ordinary skill in the art from context and understanding.

[0327] In the event that any patent, patent application, or other reference is incorporated herein by reference and (1) defines a term in a manner inconsistent with the unincorporated portion of the present disclosure or the other incorporated reference and / or (2) is otherwise inconsistent with the unincorporated portion of the present disclosure or the other incorporated reference, the unincorporated portion of the present disclosure shall control and the term or the disclosure incorporated therein shall control solely with respect to the reference in which the term was defined and / or incorporated disclosure originally existed.

[0328] Other Notes

[0329] Other embodiments according to the inventive subject matter of the present disclosure are described in the following enumerated paragraphs.

[0330] It must be noted that, as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise, such as when the word "single" is used. For example, reference to an "analyte" includes a single analyte and multiple analytes, reference to a "capture agent" includes a single capture agent and multiple capture agents, reference to a "detection agent" includes a single detection agent and multiple detection agents, and reference to a "reagent" includes a single reagent and multiple reagents.

[0331] As used herein, the terms "adapted" and "configured" mean that an element, component, or other subject matter is designed and / or intended to perform a given function. Thus, the use of the terms "adapted" and "configured" should not be interpreted as meaning that a given element, component, or other subject matter is simply "capable of" performing a given function. Similarly, a subject matter stated as being configured to perform a particular function may additionally or alternatively be described as being operable to perform that function.

[0332] As used herein, the phrase "for example", the phrase "as an example" and / or simply the terms "example" and "exemplary", when used with reference to one or more components, features, details, structures, embodiments and / or methods according to the present disclosure, are intended to convey that the described components, features, details, structures, embodiments and / or methods are illustrative, non-exclusive examples of components, features, details, structures, embodiments and / or methods according to the present disclosure. Therefore, the described components, features, details, structures, embodiments and / or methods are not intended to be limiting, required or exclusive / exhaustive; and other components, features, details, structures, embodiments and / or methods, including structurally and / or functionally similar and / or equivalent components, features, details, structures, embodiments and / or methods, are also within the scope of the present disclosure.

[0333] As used herein, the phrases "at least one" and "one or more" with respect to a list of more than one entity refer to any one or more entities in the list of entities, and are not limited to at least one of each and every entity specifically listed in the list of entities. For example, "at least one of A and B" (or equivalently, "at least one of A or B", or equivalently, "at least one of A and / or B") may refer to A alone, B alone, or A and B in combination.

[0334] As used herein, the term "and / or" placed between a first entity and a second entity means one of (1) the first entity, (2) the second entity, and (3) the first entity and the second entity. Multiple entities listed using "and / or" should be interpreted in the same manner, i.e., "one or more" of the entities so conjoined. In addition to the entities specifically identified by the "and / or" clause, other entities may optionally be present, whether related or unrelated to those entities specifically identified.

[0335] When numerical ranges are mentioned herein, the present invention includes embodiments in which the endpoints are included, embodiments in which both endpoints are excluded, and embodiments in which one endpoint is included and the other endpoint is excluded. Both endpoints should be assumed to be included unless otherwise stated. Furthermore, unless otherwise stated or apparent to one of ordinary skill in the art from context and understanding.

[0336] If any patent, patent application, or other reference is incorporated herein by reference and (1) defines a term in a manner inconsistent with the unincorporated portion of this disclosure or the other incorporated reference and / or (2) is otherwise inconsistent with the unincorporated portion of this disclosure or the other incorporated reference, the unincorporated portion of this disclosure shall control and the term or the disclosure incorporated therein shall control solely with respect to the reference in which the term was first defined and / or the disclosure originally appeared.

Claims

1. A device for measuring a thin fluid sample layer, comprising: A first plate, a second plate, a plurality of spacers, and a fixture, wherein: i. The first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration; ii. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample; iii. One or both of the plates comprise the spacer fixed to the respective plate; iv. said spacers having a predetermined substantially uniform height equal to or less than 200 microns, wherein at least one of said spacers is within said sample contact region; as well as v. the clamp compresses the first plate and the second plate to fix the two plates together in the closed configuration, wherein the pressure of the clamp inserted on the plate is 0.01 kg / cm^2 or more, wherein in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates; and wherein in a closed configuration configured after the sample is deposited in an open configuration, at least a portion of the sample is compressed by two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and regulated by the plates and the spacers.

2. A device for rapidly changing the temperature of a thin fluid sample layer, comprising: A first plate, a second plate and a spacer, wherein: i. The first plate has a thickness of 100 μm (micrometers) or less; ii. the first plate and the second plate are movable relative to each other into different configurations, including an open configuration and a closed configuration; iii. Each of the plates comprises a sample contact area on its respective surface for contacting a fluid sample; iv. one or both of the plates comprise said spacers fixed to the respective plates; v. The spacers have a predetermined substantially uniform height equal to or less than 200 microns; vi. at least one of the spacers is located in the sample contact area; wherein in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by the spacer, and the sample is deposited on one or both of the plates; and wherein in a closed configuration configured after the sample is deposited in the open configuration, at least a portion of the sample is compressed by the two plates into a layer of substantially uniform thickness and substantially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and regulated by the plates and the spacer; and Wherein the plate and the fixture are configured to allow the temperature of at least a portion of the sample to change at a rate of 10° C. / s or greater.

3. The apparatus of any preceding claim, further comprising a radiation absorbing layer located adjacent at least a portion of the sample of uniform thickness, wherein the sample and at least a portion of the radiation absorbing layer have an area substantially greater than the uniform thickness.

4. A system for rapidly changing the temperature of a thin fluid sample layer, comprising: i. A device as claimed in any preceding claim, ii. a radiation source, wherein the radiation source is configured to radiate electromagnetic waves that are significantly absorbed by the radiation absorbing layer; and iii. A controller is configured to control the radiation source and vary the temperature of the sample.

5. A method for rapidly changing the temperature of a thin fluid sample layer, comprising: i. Provide a device or system as claimed in any preceding claim; ii. depositing a fluid sample on one or both of the plates in the device; iii. after ii, pressing the plate into a closed configuration, wherein the plate compresses at least a portion of the sample into a thin layer having a thickness of less than 200 μm; as well as iv. Changing and maintaining the temperature of the sample layer by varying the presence, intensity, wavelength, frequency and / or angle of electromagnetic waves from the radiation source.

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