Device and method for examining the effect of drugs on microorganisms

Abstract

The present invention provides devices, systems and methods for performing biological and chemical assays.

Description

[0001] Cross-references

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 539,677, filed August 1, 2017, which is hereby incorporated by reference in its entirety for all purposes. Technical Field

[0003] The present invention particularly relates to apparatus and methods for conducting biological and chemical assays, especially microbiological assays. Background Art

[0004] In diagnosis and clinical settings, it is often necessary to check the impact of drugs on microorganisms. For example, sensitivity tests are usually used to help health care personnel determine which drugs or treatment strategies are most effective in treating patient infections, and sensitivity tests determine which antimicrobials will suppress the growth of bacteria or fungi that cause patient-specific infections. However, in fact, this test is usually laborious, time-consuming, and requires complicated laboratory environment and professional processing. Especially, the conventional method of usually relating to the culture of cultivating infection sites and monitoring the growth of cultures over time after using antimicrobials, the time spent in the face of frequent, rapid development or even life-threatening infections is too long. Therefore, it is highly recognized in the art that quick, easy to handle and accurate alternative tests are needed. Summary of the invention

[0005] The present invention provides an apparatus and method for solving the above-mentioned problems in examining the effects of drugs on microorganisms.

[0006] One aspect of the invention uses (a) two plates to compress a sample containing microorganisms to be examined into a thin layer, and (b), following (a), uses an imaging process to count and / or track the microorganisms over a period of time or at discrete time points.

[0007] Another aspect of the invention uses spacers to control the final sample thickness to aid in determining microbial concentration.

[0008] Another aspect of the present invention provides coating the drug to be examined on one or both plates, since the coated drug is able to dissolve and diffuse in the sample when in contact with the sample.

[0009] Another aspect of the present invention provides a multiplex capability for drug effect testing using two plates and drugs coated at different locations on the plates.

[0010] Another aspect of the present invention uses a cell viability dye which is added to the sample when the sample is loaded between two plates provided and the microbial activity is monitored with the aid of the dye.

[0011] Another aspect of the present invention provides the use of a mobile communication device (eg, a cell phone) for imaging microorganisms loaded between two provided plates.

[0012] Yet another aspect of the invention uses the mobile communication device, in addition to imaging, to notify medical personnel or other parties of test results. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] 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. Some drawings are not drawn to scale. In the drawings where experimental data points are given, the lines connecting the data points are only used to guide the observation of the data and have no other purpose.

[0014] Figure 1 An example of a general QMAX (Q: quantification; M: amplification; A: addition of reagents; X: acceleration; also known as a compression regulated open flow (CROF) device is shown.

[0015] Figure 2 An exemplary embodiment of the invention is shown in which a QMAX device is used to monitor microorganisms.

[0016] Figure 3 An exemplary embodiment of the invention is shown, in which the monitored microorganisms proliferate between two plates.

[0017] Figure 4 Another exemplary embodiment of the invention is shown, wherein the test agent affects the survival of a microorganism.

[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 subtitles used herein, if any, are for organizational purposes only and should not be construed as limiting the subject matter described in any way. The content under a section heading and / or subheading is not limited to the section heading and / or subheading, but applies to the entire description of the present invention.

[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 dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0021] Among other things, the present invention provides apparatus, systems and methods for performing biological and chemical assays using QMAX cards.

[0022] The exemplary embodiments disclosed herein may be combined with biological / chemical devices and assays, including but not limited to those disclosed, described and / or referred to in the following applications:

[0023] PCT Application No. PCT / US2016 / 045437 filed on August 10, 2016,

[0024] PCT Application No. PCT / US2016 / 051775 filed on September 14, 2016,

[0025] PCT Application No. PCT / US2016 / 051794 filed on September 14, 2016,

[0026] U.S. Provisional Application No. 62 / 369,181, filed July 31, 2016,

[0027] U.S. Provisional Application No. 62 / 412,006, filed October 24, 2016,

[0028] U.S. Provisional Application No. 62 / 437,339, filed December 21, 2016,

[0029] U.S. Provisional Application No. 62 / 431,639, filed December 9, 2016,

[0030] U.S. Provisional Application No. 62 / 456,065, filed February 7, 2017,

[0031] U.S. Provisional Application No. 62 / 456,488, filed February 8, 2017,

[0032] U.S. Provisional Application No. 62 / 456,287, filed February 8, 2017,

[0033] U.S. Provisional Application No. 62 / 456,528, filed February 8, 2017,

[0034] U.S. Provisional Application No. 62 / 456,537, filed February 8, 2017,

[0035] U.S. Provisional Application No. 62 / 456,612, filed February 8, 2017,

[0036] U.S. Provisional Application No. 62 / 456,631, filed February 8, 2017,

[0037] U.S. Provisional Application No. 62 / 456,596, filed February 8, 2017,

[0038] U.S. Provisional Application No. 62 / 456,590, filed February 8, 2017,

[0039] U.S. Provisional Application No. 62 / 456,638, filed February 8, 2017,

[0040] U.S. Provisional Application No. 62 / 456,598, filed February 8, 2017,

[0041] U.S. Provisional Application No. 62 / 456,552, filed February 8, 2017,

[0042] U.S. Provisional Application No. 62 / 456,603, filed February 8, 2017,

[0043] U.S. Provisional Application No. 62 / 456,585, filed February 8, 2017,

[0044] U.S. Provisional Application No. 62 / 456,628, filed February 8, 2017,

[0045] U.S. Provisional Application No. 62 / 456,504, filed February 8, 2017,

[0046] U.S. Provisional Application No. 62 / 456,988, filed February 9, 2017,

[0047] U.S. Provisional Application No. 62 / 457,084, filed February 9, 2017,

[0048] U.S. Provisional Application No. 62 / 457,031, filed February 9, 2017,

[0049] U.S. Provisional Application No. 62 / 456,904, filed February 9, 2017,

[0050] U.S. Provisional Application No. 62 / 457,075, filed February 9, 2017,

[0051] U.S. Provisional Application No. 62 / 457,009, filed February 9, 2017,

[0052] U.S. Provisional Application No. 62 / 457,133, filed February 9, 2017,

[0053] U.S. Provisional Application No. 62 / 457,103, filed February 9, 2017,

[0054] U.S. Provisional Application No. 62 / 459,267, filed February 15, 2017,

[0055] U.S. Provisional Application No. 62 / 459,303, filed February 15, 2017,

[0056] U.S. Provisional Application No. 62 / 459,337, filed February 15, 2017,

[0057] U.S. Provisional Application No. 62 / 459,232, filed February 15, 2017,

[0058] U.S. Provisional Application No. 62 / 459,160, filed February 15, 2017,

[0059] U.S. Provisional Application No. 62 / 459,972, filed February 16, 2017,

[0060] U.S. Provisional Application No. 62 / 394,753, filed September 15, 2016,

[0061] U.S. Provisional Application No. 62 / 459,496, filed February 15, 2017,

[0062] U.S. Provisional Application No. 62 / 459,554, filed February 15, 2017,

[0063] U.S. Provisional Application No. 62 / 460,047, filed February 16, 2017,

[0064] U.S. Provisional Application No. 62 / 459,598, filed February 15, 2017,

[0065] U.S. Provisional Application No. 62 / 460,083, filed February 16, 2017,

[0066] U.S. Provisional Application No. 62 / 460,076, filed February 16, 2017,

[0067] U.S. Provisional Application No. 62 / 460,062, filed February 16, 2017,

[0068] U.S. Provisional Application No. 62 / 459,920, filed February 16, 2016,

[0069] U.S. Provisional Application No. 62 / 459,577, filed February 15, 2017,

[0070] U.S. Provisional Application No. 62 / 459,602, filed February 15, 2017,

[0071] U.S. Provisional Application No. 62 / 460,069, filed February 16, 2017,

[0072] U.S. Provisional Application No. 62 / 460,088, filed February 16, 2017,

[0073] U.S. Provisional Application No. 62 / 460,091, filed February 16, 2017,

[0074] U.S. Provisional Application No. 62 / 460,757, filed February 18, 2017,

[0075] U.S. Provisional Application No. 62 / 463,578, filed February 24, 2017,

[0076] The entire contents of these applications are incorporated herein by reference.

[0077] The embodiments in these applications incorporated herein may be considered in combination with each other or as a single invention, rather than as discrete and independent documents. In addition, the exemplary embodiments disclosed herein are applicable to embodiments including, but not limited to, biological / chemical assays, QMAX cards and systems, QMAX with hinges, notches, grooved edges and sliders, assays and devices with uniform sample thickness, smart phone detection systems, cloud computing designs, various detection methods, labels, capture agents and detection agents, analytes, diseases, applications and samples; various embodiments are disclosed, described and / or referenced in the above applications, all of which are incorporated herein by reference in their entirety.

[0078] As used herein, the term "microorganism" refers to any organism of small size that can only be observed through a microscope, usually with an average size of less than 1 mm. Microorganisms used herein include, but are not limited to, bacteria, fungi, archaea, viruses, protozoa, and microfauna (such as myxozoa, arthropods, crustaceans, and microscopic nematodes).

[0079] The terms "drug" and "test agent" as used herein are interchangeable and refer to a biological / chemical agent whose effect on one or more microorganisms is to be examined. The term "antimicrobial agent" or "antimicrobial agent" refers to a biological / chemical agent that is capable of inhibiting the survival or growth of one or more microorganisms.

[0080] As used herein, the terms "proliferation" and "division" are interchangeable with the microorganisms as the subject thereof, and refer to the action / process taken by a microorganism individual to produce new individuals by dividing a parent individual cell into two or more daughter cells, resulting in an increase in the number of the subject microorganism individuals.

[0081] QMAX Device

[0082] Figure 1An embodiment of a general QMAX (Q: quantification); M: amplification; A: addition of reagents; X: acceleration; also known as a compression regulated open flow (CROF) device is shown. The general QMAX device comprises a first plate 10 and a second plate 2. In particular, Figure (A) shows a perspective view of the first plate 10 and the second plate 20, wherein the first plate has a spacer. However, it should be noted that the spacer can also be fixed on the second plate 20 (not shown) or on both the first plate 10 and the second plate 20 (not shown). Figure (B) shows a perspective view and a cross-sectional view of depositing a sample 90 on the first plate 10 in an open configuration. However, it should be noted that the sample 90 can also be deposited on the second plate 20 (not shown) or on both the first plate 10 and the second plate 20 (not shown). Figure (C) shows (i) the use of the first plate 10 and the second plate 20 to distribute the sample 90 (the sample flows between the inner surfaces of the plates) and reduce the sample thickness, and (ii) the use of the spacer and the plate to adjust the sample thickness in the closed configuration of the QMAX device. The inner surface of each plate has one or more binding sites and / or storage sites (not shown).

[0083] In some embodiments, the spacers 40 have a predetermined uniform height and a predetermined uniform spacer spacing. Figure 1 As shown in FIG. 4C , the spacing between the plates and thus the thickness of the sample 90 is adjusted by the spacer 40. In some embodiments, the uniform thickness of the sample 90 is substantially similar to the uniform height of the spacer 40. It should be noted that although Figure 1 A spacer 40 is shown fixed to one of the plates, but in some embodiments, the spacer is not fixed. For example, in some embodiments, the spacer is mixed with the sample so that when the sample is compressed into a thin layer, the spacer, which is a rigid bead or particle of uniform size, adjusts the thickness of the sample layer.

[0084] Microbiological monitoring

[0085] Common antimicrobial agents affect microorganisms such as bacteria or fungi by inhibiting cell proliferation processes (e.g., "bacteriostasis") or by directly killing cells (e.g., "bactericidal") by processes such as destroying the integrity of the cell wall or cell membrane. For example, penicillins and cephalosporins have bactericidal effects by targeting the bacterial cell wall, while tetracyclines are protein synthesis inhibitors that block bacterial division. Other agents can affect microorganisms by similar mechanisms or in the opposite direction, promoting the activity and / or proliferation of microorganisms. Detecting the effects of drugs on microorganisms mainly includes monitoring one or more aspects of the microorganism after the application of the test agent, such as but not limited to the number of microorganisms caused by cell proliferation or cell death, proliferation rate, cell motility, cell morphology, cell membrane permeability, and cell respiration.

[0086] One aspect of the invention is the use of the QMAX device to monitor microorganisms by one or more methods. Figure 2 An exemplary embodiment of the present invention is shown, in which the QMAX device is used to monitor microorganisms. aa' 10, a perspective view, a top view and a cross-sectional view taken along the line. The QMAX device includes a first plate 10, a second plate 20 and a spacer, and is in its closed configuration as shown. In the closed configuration, a sample 90 containing a microorganism 92 to be analyzed is confined in a thin layer by two plates. The thickness of the thin layer 90 is adjusted by the two plates and the spacer 40. In this exemplary embodiment, the thin layer thickness 910 is equal to the spacer height, and the spacer height is selected so that the microorganism 92 is confined in a single layer between the plates without being compressed. Therefore, depending on the suspected effect of the test reagent on the microorganism to be analyzed, the microorganism 92 is easy to be observed and monitored by one or more methods. Below we discuss some exemplary aspects of the microorganism to be monitored under the influence of the test reagent.

[0087] Figure 3 An exemplary embodiment of the present invention is shown, wherein monitoring microorganisms proliferate between two plates. Figure (A) and (B) respectively show the device in a closed configuration at two different time points after the sample containing microorganisms is loaded into the device. The top view of the device is shown on the left, and the cross-sectional view taken along line bb' or cc' is shown on the right. As shown in the figure, the QMAX device comprises a first plate 10, a second plate 20 and a spacer 40. Figure (A) shows the device and microorganisms shortly after the two plates enter the closed configuration, wherein the sample is limited in a thin layer by the two plates (the liquid phase component of the sample is not shown), and in this exemplary embodiment, the height of the spacer 40 is selected so that the microorganisms 92 are limited in a monolayer by the two plates. Figure (B) shows a device with microorganisms within a certain period of time after the two plates enter the closed configuration when some microorganisms have begun to split and produce new single microorganisms. For example, the microorganisms 92 represented by the dotted circle in Figure (A) and (B) produce the new individuals 94 in Figure (B). It should be noted that although depicted as stationary over time in the figures, microorganisms, particularly living individuals, between the two plates may move on their own despite the fact that in the thin layer the fluid sample is stagnant relative to the plates in the closed configuration.

[0088] Figure 4 Another exemplary embodiment of the present invention is shown, wherein the test agent affects the survival of microorganisms. A top view of the device is shown on the left, and a cross-sectional view taken along line bb', dd' or ee' is shown on the right. Figure (A) depicts a device with Figure 3Figure (A) of Figure 1 shows the same situation, while Figures (B) and (C) describe the possible situation that some microorganisms die for a certain period of time after the two plates enter the closed structure. In one case, the individual microorganism indicated by the dotted circle in Figure (A) dies, and its cell integrity is destroyed and looks "disappeared", as represented by the empty dotted circle in Figure (B). In another case, the individual microorganism shown in the dotted square in Figure (A) dies, but it maintains its cell morphology. In this case, the cell viability dye that stains the cell corpse indicates the dead individual 96 in the dotted square in Figure (C).

[0089] In some embodiments, Figure 3 In some embodiments, it is generally necessary to monitor cell proliferation, wherein the proliferation of microorganisms is suspected to be one aspect affected by the test agent. In some embodiments, it is generally necessary to monitor cell number, wherein changes in whether and how microorganisms proliferate (increase in number) and / or die-disappear (decrease in number) are observed under the influence of the test agent, respectively, as shown in FIG. Figure 3 and Figure 4 (B) As shown. In some embodiments, it is necessary to monitor cell activity, wherein it is suspected that under the influence of the test agent, it is observed whether and how the microorganism dies but maintains its cell morphology, such as Figure 4 Group (C) shown.

[0090] However, in some embodiments, it is desirable to monitor cell morphology, where it is suspected that their morphology is an aspect affected by the test reagent. In some embodiments, it is desirable to monitor cell motility, where it is suspected that cell motility is an aspect affected by the test reagent. In some embodiments, it is desirable to monitor some other aspect of the microorganisms as other needs may be met.

[0091] In some embodiments, it may be desirable to monitor any combination of the foregoing aspects of a microorganism.

[0092] An important aspect of the present invention is that the QMAX device is capable of implementing a different range of monitoring possibilities to meet different needs, as will be described in further detail below.

[0093] Reagents on the plate

[0094] In order to examine the effects of drugs on microorganisms, it is often necessary to contact the microorganisms with various reagents, including test reagents. In some embodiments, the various reagents are added to the sample containing the microorganisms before the sample is deposited on one or both plates. However, one aspect of the present invention is to provide QMAX devices and methods in which the reagents are coated on one or both plates and added to the sample when contacting the sample.

[0095] In some embodiments, the sample to be analyzed by the QMAX device is preloaded with a test reagent, and the QMAX device is used only to monitor the microorganisms over time. In some embodiments, the test reagent is coated on the sample contact area of ​​one or both plates and is configured to dissolve into and diffuse in the sample upon contact with the sample. In some embodiments, the test reagent comprises at least two parts, one of which is preloaded into the sample and the other is coated on the plate.

[0096] In some embodiments, multiplexed testing is performed using a single QMAX device. In some embodiments, different types of test reagents are applied at different locations in the sample contact area of ​​the plate. In some embodiments, the same type but different concentrations of test reagents are applied at different locations in the sample contact area of ​​the plate. In some embodiments, different test reagents and / or different concentrations of the same test reagent are applied to the plate by fluid isolation, for example, there is a wall structure on the plate sample contact area to isolate different sub-areas. In some embodiments, there is no fluid isolation between different sub-areas, and due to limited lateral diffusion in the thin layer, the sub-areas are "spaced" from each other.

[0097] In certain embodiments, one or two on the plate comprise a control area and an experimental area on the corresponding sample contact area, wherein the experimental area comprises one or more test reagents, and the test reagent dissolves and diffuses in the sample when contacting the sample, and wherein the control area does not comprise this type of test reagent. In certain embodiments, the control area and the experimental area are fluid-isolated. In certain embodiments, the control area and the experimental area are not fluid-isolated, and microorganisms can not move through different districts in the time period of inspection. Comparing the microorganisms in the experimental area and the control area is conducive to measuring the impact of the test reagent on the microorganism.

[0098] In some embodiments, the test agent includes an antibiotic, such as, but not limited to, actinomycin D, actinomycin, aculactin A, acyclovir, adenine 9-β-D-arabinofuranoside, alamethicin, alamethicin, L-alanine-L-1-aminoethylphosphonic acid, albendazole, 17-(allylamino)-17-demethoxygeldanamycin, astatin, amikacin, amikacin, 7-aminoactinomycin D, 7-aminocephalosporanic acid, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, (+)-6-aminopenicillanic acid, amoxicillin, amphotericin B, ampicillin, anhydroerythromycin A, anisomycin, antimycin A, antimycin A2, antipain, aphidicolin, aphidicolin, apoptolidin A, Apramycin, Artesunate, Ascochlorin, Ascomycin, 5-Azacytidine, Azaserine, Azithromycin, Azlocillin, Bacitracin, Antimicrobial Peptides, Bafilomycin A1, Bafilomycin B1, Benstatin, βD-4 Human Recombinant, βD-1 (36 amino acids) Human Recombinant, βD-1 (47 amino acids) Human Recombinant, Bithionol VETRANAL TM , Blasticidin S, Bleomycin, BM-Cyclin, Borrelidin, Brefeldin A, Penicillin A, Calcium Ionophore III Selector TM , calcium ionophore A23187, calcium ionophore A23187, calcium ionophore A23187, (S)-(+)-camptothecin, capreomycin, carbadox, carbenicillin, carboplatin, cecropin A, cecropin B, cecropin P1, cefaclor, cephalexin VETRANAL TM, cefazolin, cefixime, cefmetazole, cefoperazone, cefotaxime, cefsulodin, ceftazidime, ceftriaxone, cephalexin, cephalosporin, cefotaxime, cephradine, cercosporin, cerulenin, cetylpyridine, chloramphenicol, chlorhexidine, chloroquine, chlortetracycline, chromomycin A3, chrysomelin A, chrysomelin B, cinnamycin, cinoxacin, ciprofloxacin, clarithromycin, clindamycin, clofazimine, clotrimazole, o-cloxacillin, colistin, polyacetylneuraminic acid, compound 48 / 80, concanavalin A, cordycepin, kumarin A1, cryptotanshinone, crystal violet, cyclo Hexamidine, D-cycloserine, cyclosporin A, cyclosporin C, cytochalasin D, cytochalasin B, dacarbazine antitumor purine analog, daunorubicin, 10-deacetylbaccatin III, dequamectin, defensin HNP-1, defensin HNP-2, demeclocycline hydrochloride, 1-deoxymannonojirimycin, 1-deoxynojirimycin, cis-diammineplatinum (II), 6,9-diamino-2-ethoxyacridine-DL-lactate, cis-diammineplatinum (II), 6-diazo-5-oxo-L-norleucine, 5,7-dichloro-8-hydroxy-2-methylquinoline, dichlorophenol Dicloxacillin, diethylcarbamazine, diethylcarbamazine, difloxacin, dihydrostreptomycin, dihydrostreptomycin, diloxanide furoate, dimethoprim, dirithromycin, doxorubicin, doxycycline, duramycin, econazole, elastin, anthracene, emetine, enrofloxacin, erythromycin, ethambutol, etoposide, fengyuansu, serine, florfenicol, flubendazole VETRANAL TM, Fluconazole, Flumequine, Flumethasone, 5-fluorocytidine nucleoside analogs, Flurbiprofen cyclooxygenase inhibitors, Metamycin A, Fumagillin, Fumagillin C, Furazolidone, Fusaric acid, G 418, Ganciclovir, Gatifloxacin, Geldamicin, Gentamycin, Gentamycin, Crystal violet, Gliotoxin, Gramicidins, Griseofulvin, Herbimycin A, Cetylpyridine, Magnolia nobile, Hydrocortisone 21, 8-Hydroxyquinoline, 4-Hydroxytamoxifen, (Z)-4-Hydroxytamoxifen, Hygromycin B, Cantharicin, Imipenem, Indomethacin, Indomethacin, Ionomycin, Ir gasan, itraconazole, iturin A, ivermectin, josamycin, K-252a, K-252b, kanamycin, kasugamycin, kendomycin, ketoconazole, levomycin, L-(+)-lactic acid, lactoferrin B, leptomycin A, leptomycin B, levamisole, levofloxacin, lincomycin, Listeria monocytogenes selective supplement I, LL-37, lomefloxacin, lysostaphin, bombesin I, mebendazole, meclocycline, menadione, 2-thiopyridine N-oxide, N-methyl-1-deoxynojirimycin, 2-methyl-4-isothiazoline-3-hydrochloride, metronidazole, mevastatin, (±)-miconazole, minocycline, mithramycin A, mitomycin C, monensin, morantel, hydroxycarboxylic acid, mupirocin, subtilisin, myxothiazole, propylthioamide, nafcillin, naftifine, nalidixic acid, methyl salt mycin, neocarcinostatisin, neomycin, netilmicin, neterotene, niclosamide, nigericin, nikkomycin Z, nisin, nitrofurantoin, nogamycin, no active mycin, norfloxacin, nourseothricin, neomycin, NP-1, nystatin, ochratoxin A, ofloxacin, oligomycin, oligomycin A, neocyandiamide, octacycline, oxazolidinone, oxytetracycline, oxytetracycline, paclitaxel, paromomycin, patulin, PD 404,182, pediocin, pefloxacin, D-penicillamine, penicillin G, penicillin V, pentamidine, PGLa, 1,10-phenanthroline, phenazine, penicillin V, phleomycin, fosfomycin, pimaricin, pipemidic acid, piperacillin, pirarubicin, plate mycin, polymyxin B, poly(vinyl pyrrolidone) iodine complex, potassium clavulanate:cellulose (1:1), potassium sorbate, praziquantel, puromycin, pyrantel, pyrazinamide, bisquinacridone, pyrrolocin, quinine, 8-quinolinol, radicicol, ramoplanin, rapamycin, butterfly mycin, rafamycin A, ribavirin, ribostamycin, albendazole sulfoxide, rifabutin, rifampicin, rifamycin SV, rifapentine, rifaximin, ristocetin, rolicycline, roxithromycin , Salinomycin, Sangmycin, Sinaifungin, Perillomycin, Sorbic acid, Sordolin, Sparfloxacin, Spectinomycin, Spergualin, Spiramycin, Staurosporine, Streptolysin O, Streptomycin, Streptomycin, Streptomycin Melanosporin, Streptozotocin, Succinylsulfathiazole, Sulconazole, Sulfadiazine, Sulfadiazine, Sulfadiazine, Sulfadimethoxine, Sulfadimethoxine, Sulfadiaz ... E, tamoxifen, tazobactam, teicoplanin, terbinafine, terconazole, tetracycline, tetramidazole, thiabendazole, thiamphenicol, sodium thimerosal, thiocandin, thiostrepton, thio-TEPA, thymol, tiamulin, hydroxythiophene penicillin, tioconazole, tobramycin, aminoglycoside antibiotics, tobramycin, tolnaftate, toyocamycin, triazine C, trichlorfon, trimethoprim, tuberculin, tunicamycin, tunicamycin C2 homologue, tylosin, valacyclovir, valinomycin, vinblastine, virginiamycin S1, virginiamycin M1, and any analogs, salts and derivatives thereof. ,

[0099] In some embodiments, the test agent includes an antifungal agent such as, but not limited to, a polyene antifungal agent (amphotericin B, candidiasis, filipinocin, harstomycin, natamycin, nystatin, chelamycin), an imidazole (e.g., bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole), a triazole (albaconazole, epoxiconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, ravuconazole, terconazole, voriconazole), thiazoles (e.g., abafungin), allylamines (e.g., amorolfine, butenafine, naftifon and terbinafine), echinocandins (e.g., anidulafungin, caspofungin and micafungin), aurone, benzoic acid, ciclopirox, flucytosine and 5-flucytosine, griseofulvin, chloropropynyl iodide, tolnaftate, undecylenic acid, crystal violet, Peruvian balsam, analogs thereof, salts thereof and derivatives thereof.

[0100] In addition to or in place of test reagents, in some embodiments, the QMAX device includes on one or both plates various other reagents to be added to the sample, such as, but not limited to, staining dyes to label microorganisms (cell viability dyes as well as many others), life support reagents to promote microbial survival (e.g., nutrients, oxygen sources, pH buffers), proliferation reagents to maintain or promote microbial proliferation (e.g., hormones, siderophores).

[0101] In some embodiments, cell viability dyes include, but are not limited to, propidium iodide (PI), 7-AAD (7-aminoactinomycin D), trypan blue, calcein violet AM, calcein AM, fixable viability dyes (FVD) conjugated to different fluorophores, SYTO9 and other nucleic acid dyes, resazurin and colored formazan (MTT / XTT) and other mitochondrial dyes, and any combination thereof, etc. In some embodiments, the sample contains bacteria, and it is necessary to determine the viability of bacteria in the sample, and the device also contains bacterial viability dyes, such as PI, SYTO9, etc., on one or both plates to differentially stain live cells from dead cells.

[0102] In some embodiments, the dye is selected from the group consisting of acid fuchsin, alcian blue 8GX, alizarin red S, aniline blue WS, auramine O, azo carmine B, azo carmine G, sky blue A, sky blue B, sky blue C, basic fuchsin, Bismarck brown Y, brilliant cresyl blue, brilliant green, carmine, chlorazole black E, Congo red, CI cresyl violet, crystal violet, Darrow red, eosin B, eosin Y, erythrosine, ethyl eosin, ethyl green, fast green FCF, fluorescein isothiocyanate, Giemsa dye, hematoxylin, hematoxylin & eosin, indigo carmine, Janus Green B, MacNeal's dye 1899, Light Green SF, Malachite Green, Matthew's Yellow, Methyl Orange, Methyl Violet 2B, Methylene Blue, Methylene Blue, Methylene Violet, (Methylene Blue), Neutral Red, Aniline Black, Nile Blue A, Nuclear Fast Red, Oil Red, Orange G, Orange II, Orcein Red, Parafuchsin, Bamboo Red B, Strong Protein Silver S, Pyronine B, Pyronine, Resazurin, Rose Bengal, Safranin O, Sudan Black B, Sudan III, Sudan IV, Tetrachrome Stain (MacNeal), Lowe's Violet, Toluidine Blue, Weigert, Wright's Stain, and any combination thereof.

[0103] In some embodiments, the dye is conjugated to a fluorescent molecule (fluorophore), including but not limited to, IRDye800CW, Alexa 790, Dylight 800, fluorescein, fluorescein isothiocyanate, succinimidyl ester of carboxyfluorescein, succinimidyl ester of fluorescein, 5-isomer of fluorescein dichlorotriazine, caged carboxyfluorescein-alanine-carboxamide, Oregon Green 488, Oregon Green 514; fluorescein yellow, acridine orange, rhodamine, tetramethylrhodamine, Texas Red, propidium iodide, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide), tetrabromorhodamine 123, rhodamine 6G, TMRM (tetramethylrhodamine methyl ester), TMRE (tetramethylrhodamine ethyl ester), tetramethylrhodamine, rhodamine B and 4-dimethylaminotetramethylrhodamine, Green chromatic fluorescent protein, blue-shifted green fluorescent protein, blue-green-shifted green fluorescent protein, red-shifted green fluorescent protein, yellow-shifted green fluorescent protein, 4-acetamido-4'-isothiocyanate stilbene-2,2'-disulfonic acid; acridine and derivatives, for example, acridine, acridine isothiocyanate; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS); 4-amino-N-[3-vinylsulfonyl)phenyl]naphthalenamide-3,5-disulfonate; N-(4-anilino-1-naphthyl)maleimide; anthranilamide; 4,4-difluoro-5-(2-thienyl)-4-boron-3a,4adiaza-5-indacene-3-propionic acid BODIPY; Cascade Blue; Brilliant Yellow; Coumarin and derivatives: Coumarin, 7-amino-4-methylcoumarin (AMC, coumarin 120), 7-amino-4-methylcoumarin (coumarin 151); cyanine dyes; label red (cyanosine); 4',6-diamino-2-phenylindole (DAPI); 5',5"-dibromopyrogallolsulfonaphthalene (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; diethylenetriamine pentaacetate; 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid; 4,4'-diisothiocyanato-2,2'-yldisulfonic acid disodium salt; 5-(dimethylamino)naphthalene-1-sulfonyl chloride (DNS, dansyl) chlorine); 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC); eosin and derivatives: eosin, eosin isothiocyanate, erythrophyca and derivatives: erythrophyca B, erythrophyca, isothiocyanate; ethidium; fluorescein and its derivatives: 5-carboxyfluorescein (FAM), 5-(4,6-dichlorotriazin-2-yl)amino-fluorescein (DTAF), 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein (JOE), fluorescein, fluorescein isothiocyanate, QFITC, (XRITC); fluorescamine; IR144; IR1446; malachite green isothiocyanate; 4-methylumbelliferyl o-cresolphthalein; nitrotyrosine; parafuchsin; phenol red;B-phycoerythrin; o-phthalaldehyde; pyrene and its derivatives: pyrene, pyrenebutyric acid, succinimidyl 1-pyrene; butyrate quantum dots; Reactive Red 4 (Cibacron; TM Brilliant Red 3B-A) Rhodamine and derivatives: 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), lissamine rhodamine B sulfonyl chloride rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine X isothiocyanate, sulforhodamine B, sulforhodamine 101, sulfonyl chloride derivatives of 5-sulforhodamine (Texas Red); N,N,N',N'-tetramethyl-6-carboxyrhodamine TAMRA; tetramethylrhodamine; tetramethylrhodamine isothiocyanate (TRITC); riboflavin; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS), 4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL), rhodamine; CAL fluorescent orange 560; terbium chelate derivatives; Cy3; Cy5; Cy5.5; Cy7; IRD700; IRD 800; La Jolla Blue; phthalocyanines; and naphthalocyanines, coumarins and related dyes, xanthene dyes such as rhodols, resorufins, bimanes, acridines, isoindoles, dansyl dyes, aminophthalhydrazides (e.g., luminol), and isoluminol derivatives, aminophthalimides, aminonaphthalimides, aminobenzofurans, aminoquinosides, dicyanohydroquinones, fluorescent europium and terbium complexes; combinations thereof, and the like. Suitable fluorescent and chromogenic proteins include, but are not limited to, green fluorescent protein (GFP), including, but not limited to, GFP derived from Aequoria victoria or a derivative thereof, e.g., a "humanized" derivative, such as enhanced GFP; GFP from another species, such as Renilla reniformis, Renilla mulleri, or Ptilosarcus guernyi; "humanized" recombinant GFP (hrGFP); any of a variety of fluorescent and colored proteins from coral species; any combination thereof; and the like.

[0104] Imaging and Image Processing

[0105] In some embodiments, examination of the effect of a drug on a microorganism is accomplished by monitoring the microorganism in a uniform thickness layer and determining the effect of the test agent on the microorganism when the two plates are in a closed configuration as described above.

[0106] In some embodiments, monitoring is accomplished optically, and one or both of the plates are transparent, enabling optical inspection and / or imaging of microorganisms between the two plates.

[0107] In some embodiments, microscopy techniques are used to monitor microorganisms in the device. In some embodiments, a standard laboratory microscope is used, while in some embodiments, a miniaturized or custom designed microscope is used.

[0108] In some embodiments, monitoring is accomplished by imaging. An imager is used to obtain images of signals emitted from microorganisms in the device. In some embodiments, the imager comprises a camera and a light source for imaging the microorganisms in the thin layer.

[0109] In some embodiments, a series of images are taken of the microorganisms in the device at discrete time points. In some embodiments, images of the microorganisms in the device are taken over a period of time.

[0110] In some embodiments, visual inspection of the microorganisms is performed by a professional or non-professional user of the device with the aid of a microscope, an acquired image of the microorganisms, and / or an acquired video.

[0111] In some embodiments, the processor is used to process the obtained images of microorganisms and / or the obtained videos. The processor includes electronics, a signal processor, and hardware and software for receiving and processing images and identifying and analyzing platelets in the images.

[0112] In some embodiments, the monitoring and determining steps comprise:

[0113] i. obtaining a series of images of a plate containing microorganisms in a sample layer of uniform thickness between the plates at discrete time points;

[0114] ii. identifying and / or counting microorganisms in at least one region of each image; and

[0115] iii. Assessing changes in the microorganisms between at least some discrete time points.

[0116] In some embodiments, the monitoring and determining steps comprise:

[0117] i. acquiring a video comprising a series of images of a plate at discrete time points during a first time period, the plate containing microorganisms in a layer of uniform thickness;

[0118] ii. identifying and tracking at least a portion of the microorganisms in the video; and

[0119] iii. evaluating changes in the tracked microorganism during at least a portion of the first time period.

[0120] In some embodiments, the identifying step comprises identifying a microorganism stained with a dye, and wherein the dye comprises a cell viability dye that indicates whether the stained cell is alive or dead.

[0121] In some embodiments, the changes in microorganisms include, but are not limited to, changes in the total number of microorganism individuals, changes in microorganism concentration, changes in microorganism morphology, changes in microorganism motility, changes in microorganism absorption of staining dyes, and any combination thereof.

[0122] In some embodiments, changes in the number or concentration of individual microorganisms are evaluated. The processor is configured to count the number of identified microorganisms in at least one region of each image. In some cases, the concentration of the microorganisms is determined. An advantage of the present invention is that in some embodiments, the spacer controls a very uniform thickness of the final sample layer. Therefore, by multiplying the region of interest by the final sample thickness, the volume of the sample in at least one region of each image can be easily calculated, and the concentration of the microorganisms in the region of interest can be easily derived.

[0123] In some embodiments, the change in the number of individual microorganisms is evaluated by tracking at least a portion of the microorganisms in the obtained video. When tracking individual microorganisms, the processor is also configured to identify events when the tracked individuals undergo cell division (producing new individuals and increasing in number) and / or disappear due to cell death and cell body decomposition.

[0124] In some embodiments, changes in microbial morphology are assessed by analyzing geometric parameters of images of each individual microorganism in the acquired images and / or videos.

[0125] In some embodiments, changes in microbial motility are assessed by analyzing movement parameters of tracked microbial individuals in acquired videos, such as, but not limited to, movement speed, direction, and duration and frequency of immobility.

[0126] In some embodiments, changes in the uptake of a staining dye by a microorganism are assessed by evaluating the signal emitted by the staining dye within the boundaries of individual microorganisms.

[0127] System for examining the effects of drugs on microorganisms

[0128] Another aspect of the present invention is to provide a system for examining the effects of drugs on microorganisms. And in certain embodiments, the system allows remote health monitoring, consultation, etc.

[0129] In some embodiments, the system includes:

[0130] (a) QMAX device;

[0131] (b) an imager comprising a camera and a light source for imaging microorganisms in a layer of uniform thickness; and

[0132] (c) A processor comprising electronics, a signal processor, and hardware and software for receiving and processing images and identifying and analyzing microorganisms in the images.

[0133] In some embodiments, the imager is configured to obtain a series of images of the microorganism at discrete points in time.

[0134] In some embodiments, the processor is configured to receive and analyze the series of images, identifying and counting microorganisms in a first region of each image.

[0135] In some embodiments, the imager is configured to acquire a video comprising a series of images of the microorganisms over a specific period of time.

[0136] In some embodiments, the processor is configured to receive and analyze the video, identify and track at least a portion of the microorganisms in the video, and evaluate changes in the tracked microorganisms over at least a portion of the first time period.

[0137] In some embodiments, the mobile communication device is used as an imager and optionally as an image processor. In some embodiments, the system includes:

[0138] (a) A QMAX device as described in any preceding or following embodiment;

[0139] (b) A mobile communication device comprising:

[0140] i. an imager comprising one or more cameras for imaging microorganisms in a sample;

[0141] ii. a processor comprising electronics, a signal processor, and hardware and software for receiving and / or processing images of microorganisms and for remote communication; and

[0142] (c) a light source from the mobile communication device or an external source, wherein the light source is configured to provide illumination to the sample for imaging with the camera.

[0143] In some embodiments, the system further comprises:

[0144] (d) A housing configured to hold a sample and to be mounted to a mobile communication device.

[0145] In some embodiments, the housing includes optics for facilitating imaging and / or signal processing of a sample by a mobile communication device, and a mount configured to hold the optics on the mobile communication device.

[0146] In some embodiments, the mobile communication device is configured to transmit the test results to medical personnel, a medical institution, or an insurance company.

[0147] In some embodiments, the mobile communication device is further configured to communicate information about the subject with medical personnel, medical institutions, or insurance companies. In some embodiments, the mobile communication device is configured to receive prescriptions, diagnoses, or recommendations from medical personnel. In some embodiments, the mobile communication device communicates with a remote location via WiFi or a cellular network.

[0148] In some embodiments, the mobile communication device is a mobile phone.

[0149] application

[0150] The present invention has particular use in many aspects of a clinical / diagnostic setting.

[0151] In some embodiments, the present invention can be used for susceptibility testing, wherein the effect of antimicrobial drugs on potential microbial pathogens causing patient infections is screened to help medical personnel select the most effective antimicrobial drugs or treatment strategies for treating patient infections. In some embodiments, testing is performed to facilitate identification of pathogenic microorganisms. In some embodiments, QMAX devices, systems and methods are used to examine the effect of antimicrobial drugs on potential microorganisms causing patient or other subject conditions. In some embodiments, high throughput testing of various antimicrobial drugs at various concentrations / doses is applied in parallel to potential microbial pathogens using a single device or system configured with multiplexing functionality as described above.

[0152] Table 1 is a chart of common pathogens. In some embodiments, the present invention can be used to test the effect of a drug on one or more of the bacteria listed in the chart.

[0153] Table 1 Common pathogens

[0154]

[0155]

[0156]

[0157] In certain embodiments, the present invention can be used for many other tests related to biological and / or clinical samples except sensitivity test.These samples include but are not limited to cells, tissues and body fluids.Interested body fluids include but are not limited to amniotic fluid, aqueous humor, vitreous humor, blood (e.g., whole blood, fractionated blood, plasma, serum), breast milk, cerebrospinal fluid (CSF), cerumen (ear wax), chyle, chyme, endolymph, perilymph, excreta, gastric acid, gastric juice, lymph, mucus (including nasal drainage and sputum), pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatism, saliva, sebum (skin oil), semen, sputum, sweat, synovial fluid, tears, vomitus, urine and exhaled condensate.In certain embodiments, the sample comprises human body fluid. In some embodiments, the sample comprises at least one of: cells, tissues, body fluids, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, cerumen, chyle, chyme, endolymph, perilymph, feces, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine, and exhaled condensate.

[0158] In some embodiments, the present invention can be used for many other tests involving microorganisms, such as but not limited to environmental testing, food testing, and forensic testing. Environmental samples can be obtained from any suitable source, such as rivers, oceans, lakes, rainwater, snow, sewage, sewage treatment runoff, agricultural runoff, industrial runoff, tap water or drinking water, etc. Food samples can be obtained from any suitable source, such as tap water, drinking water, prepared foods, processed foods, or unprocessed foods, etc.

[0159] In some embodiments, the sample is obtained directly from a source. In some embodiments, the sample is pre-loaded with reagents or pre-treated according to various needs before being deposited on a plate of the device. In some embodiments, when it is necessary to increase the level of the microorganism to be analyzed, the sample to be deposited on the device comprises a product from a sample culture obtained directly from a source.

[0160] Control plate spacing and sample thickness using spacers

[0161] According to the invention, the spacing between the two plates and thus the sample thickness is controlled by using spacers.

[0162] Spacer height. In some embodiments, all spacers have the same predetermined height. In some embodiments, the spacers have different predetermined heights. In some embodiments, the spacers can be divided into groups or regions, where each group or region has its own spacer height. And in some embodiments, the predetermined height of the spacers is the average height of the spacers. In some embodiments, the heights of the spacers are approximately the same. In some embodiments, a percentage of the spacers have the same height.

[0163] 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.

[0164] 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.

[0165] In some embodiments, the spacer height is precisely controlled. The relative accuracy of the spacers (i.e., the ratio of the deviation to the desired spacer height) is 0.001% or less, 0.01% or less, 0.1% or less; 0.5% or less, 1% or less, 2% or less, 5% or less, 8% or less, 10% or less, 15% or less, 20% or less, 30% or less, 40% or less, 50% or less, 60% or less, 70% or less, 80% or less, 90% or less, 99.9% or less, or in a range between any two of the values.

[0166] In some embodiments, the spacer height, spacing between plates, and / or sample thickness is: (i) equal to or slightly larger than the smallest dimension of the analyte, or (ii) equal to or slightly larger than the largest dimension of the analyte. "Slightly larger" means about 1% to 5% larger, and any value between the two values.

[0167] In some embodiments, the spacer height, spacing between plates, and / or sample thickness is greater than the smallest dimension of the analyte (eg, the analyte has an anisotropic shape), but less than the largest dimension of the analyte.

[0168] For example, a red blood cell has a disc shape with a minimum dimension of 2 μm (disc thickness) and a maximum dimension of 11 μm (disc diameter). In an embodiment of the present invention, the spacer is selected so that the inner surface spacing of the plate in the relevant area is 2 μm (equal to the minimum dimension) in one embodiment, 2.2 μm in another embodiment, or 3 μm (50% larger than the minimum dimension) in another embodiment, but less than the maximum dimension of the red blood cell. Such an embodiment has certain advantages in blood cell counting. In one embodiment, for red blood cell counting, by making the inner surface spacing 2 μm or 3 μm and any numerical value between the two values, an undiluted whole blood sample is confined to this spacing; on average, each red blood cell (RBC) does not overlap with others, thereby allowing the red blood cells to be accurately counted visually. (Too much overlap between RBCs may cause serious errors in counting).

[0169] In some embodiments, the spacer height, spacing between plates, and / or sample thickness is: (i) equal to or slightly smaller than the smallest dimension of the analyte, or (ii) equal to or slightly smaller than the largest dimension of the analyte. "Slightly smaller" means that it is about 1% to 5% smaller, and any number between the two values.

[0170] In some embodiments, the spacer height, spacing between plates, and / or sample thickness is greater than the smallest dimension of the analyte (eg, the analyte has an anisotropic shape), but less than the largest dimension of the analyte.

[0171] In the present invention, in some embodiments, the plates and spacers are used not only to adjust the thickness of the sample, but also to adjust the orientation and / or surface density of the analytes / entities in the sample when the plate is in a closed configuration. When the plate is in a closed configuration, a thinner thickness of the sample results in fewer analytes / entities per surface area (i.e., a smaller surface concentration).

[0172] Spacer Transverse Dimensions. For open spacers, the transverse dimensions can be characterized by their transverse dimensions (sometimes referred to as widths) in two orthogonal directions, x and y. The transverse dimensions of the spacer in each direction may be the same or different. In some embodiments, the lateral dimension in each direction (x or y) is 1 nm or less, 3 nm or less, 5 nm or less, 7 nm or less, 10 nm or less, 20 nm or less, 30 nm or less, 40 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, or 500 μm or less, or in a range between any two of said values.

[0173] In some embodiments, the ratio of the lateral dimensions in the x and y directions is 1, 1.5, 2, 5, 10, 100, 500, 1000, 10,000, or in the range between any two values. In some embodiments, different ratios are used to adjust the sample flow direction; the larger the ratio, the flow is along one direction (the larger dimension direction).

[0174] In some embodiments, the different lateral dimensions of the spacers in the x and y directions serve to (a) use the spacers as scale markers to indicate plate orientation, (b) use the spacers to create more sample flow in a preferred direction, or both.

[0175] In a preferred embodiment, the period, width and height of the spacers are substantially the same. In some embodiments, all spacers have the same shape and size. In some embodiments, the spacers have different lateral dimensions.

[0176] For the enclosing spacer, in some embodiments, the inner lateral shape and dimensions are selected based on the total volume of the sample to be enclosed by the enclosing spacer, where the volume dimensions have been described in the present disclosure; and in certain embodiments, the outer side shape and dimensions are selected based on the required strength to support the pressure of the liquid against the spacer and the compression pressure of the press plate.

[0177] In certain embodiments, the aspect ratio of the height to the average lateral dimension of the column spacers is 100,000, 10,000, 1,000, 100, 10, 1, 0.1, 0.01, 0.001, 0.0001, 0,00001, or a range between any two of the aforementioned values.

[0178] Spacer spacing. The spacer can be a single spacer or a plurality of spacers on the plate or in the sample-related area. In some embodiments, the spacers on the plate are configured and / or arranged in an array, and the array is fixed spacing, a non-fixed spacing array, or fixed spacing at some locations on the plate and non-fixed spacing at other locations.

[0179] In some embodiments, the spaced array of spacers is arranged in a lattice of squares, rectangles, triangles, hexagons, polygons, or any combination thereof, where combination means that different locations of the plate have different spacer grids.

[0180] In some embodiments, the spacer spacing of the spacer array is fixedly spaced in at least one direction of the array (ie, uniform spacer spacing). In some embodiments, the spacer spacing is configured to improve uniformity between plate spacings in a closed configuration.

[0181] In some embodiments, the distance between adjacent spacers (i.e., the spacer pitch) is 1 μm or less, 5 μm or less, 7 μm or less, 10 μm or less, 20 μm or less, 30 μm or less, 40 μm or less, 50 μm or less, 60 μm or less, 70 μm or less, 80 μm or less, 90 μm or less, 100 μm or less, 200 μm or less, 300 μm or less, 400 μm or less, or in a range between any two of said values.

[0182] In certain embodiments, the spacer spacing is 400 μm or less, 500 μm or less, 1 mm or less, 2 mm or less, 3 mm or less, 5 mm or less, 7 mm or less, 10 mm or less, or any range between these values. In certain embodiments, the spacer spacing is 10 mm or less, 20 mm or less, 30 mm or less, 50 mm or less, 70 mm or less, 100 mm or less, or any range between the values.

[0183] The distance between adjacent spacers (i.e., the spacer spacing) is selected so that for given properties of the plate and the sample, in the closed configuration of the plate, in some embodiments, the sample thickness between two adjacent spacers varies by at most 0.5%, 1%, 5%, 10%, 20%, 30%, 50%, 80%, or within any range between the stated values; or in certain embodiments, at most 80%, 100%, 200%, 400%, or within a range between any two of the stated values.

[0184] Obviously, in order to maintain a given sample thickness variation between two adjacent spacers, a closer spacer spacing is required when using a more flexible plate.

[0185] In a preferred embodiment, the spacers are a square array of fixed spacing, wherein the spacers are pillars with a height of 2-4 μm, an average lateral dimension of 1-20 μm, and a spacer pitch of 1 μm-100 μm.

[0186] In a preferred embodiment, the spacers are a regularly spaced square array, wherein the spacers are pillars with a height of 2-4 μm, an average lateral dimension of 1-20 μm, and a spacer pitch of 100 μm-250 μm.

[0187] In a preferred embodiment, the spacers are a regularly spaced square array, wherein the spacers are pillars with a height of 4-50 μm, an average lateral dimension of 1-20 μm, and a spacer pitch of 1 μm-100 μm.

[0188] In a preferred embodiment, the spacers are a regularly spaced square array, wherein the spacers are pillars with a height of 4-50 μm, an average lateral dimension of 1-20 μm, and a spacer pitch of 100 μm-250 μm.

[0189] The spacing of the spacer array is 1nm to 100nm in one preferred embodiment, 100nm to 500nm in another preferred embodiment, 500nm to 1000nm in a separate preferred embodiment, 1μm (i.e., 1000nm) to 2μm in another preferred embodiment, 2μm to 3μm in a separate preferred embodiment, 3μm to 5μm in another preferred embodiment, 5μm to 10μm in a separate preferred embodiment, 10μm to 50μm in another preferred embodiment, 50μm to 100μm in a separate preferred embodiment, 100μm to 175μm in a separate preferred embodiment, and 175μm to 300μm in a separate preferred embodiment.

[0190] Spacer density. The spacers are arranged on the corresponding plates at the following surface density: 2 Greater than 1, every 10μm 2 Greater than 1, every 100 μm 2 Greater than 1, every 500μm 2 Greater than 1, every 1000μm 2 Greater than 1, every 5000μm 2 Greater than 1, every 0.01mm 2 Greater than 1, every 0.1mm 2 Greater than 1, every 1mm 2 Greater than 1, every 5mm 2 Greater than 1, every 10mm 2 Greater than 1, every 100mm 2 Greater than 1, every 1000mm 2 Greater than 1, every 10000mm2 Greater than 1, or within the range between any two values. In some embodiments, the spacer has at least 1 / mm 2 , at least 10 / mm 2 , at least 50 / mm 2 , at least 100 / mm 2 , at least 1,000 / mm 2 or at least 10,000 / mm 2 density.

[0191] The spacer area fill factor is defined as the ratio of the spacer area to the total area or the ratio of the spacer spacing to the width. In some embodiments, the fill factor is at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, or in a range between any two of the values. In certain embodiments, the fill factor is at least 2.3%.

[0192] In a device containing two plates and a spacer, 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 5×10 6 μm 3 / GPa or less.

[0193] In a device containing two plates and a spacer, 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 5×10 5 μm 3 / GPa or less.

[0194] In a device comprising two plates and spacers, 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 filling factor of the spacer is equal to or greater than 2 MPa, wherein the filling factor is the ratio of the spacer contact area to the total plate area, and wherein for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1 (one).

[0195] In a device comprising two plates and spacers, 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 a size of the analyte, wherein a Young's modulus of the spacer multiplied by a packing factor of the spacer is equal to or greater than 2 MPa, wherein the packing factor is a ratio of a spacer contact area to a total plate area, and wherein for each spacer, a ratio of a lateral dimension of the spacer to its height is at least 1 (one), wherein the fourth power of the spacer spacing (ISD) divided by a thickness (h) of the flexible plate and a Young's modulus (E) (ISD) 4 / (hE)) is 5x10 6 μm 3 / GPa or less.

[0196] In a device including two plates and spacers, a ratio of a spacer pitch of the spacers to an average width of the spacers is 2 or greater, and a filling factor of the spacers multiplied by a Young's modulus of the spacers is 2 MPa or greater.

[0197] Examples of the Invention

[0198] A1. A device for examining the effect of a drug on a microorganism, comprising:

[0199] A first plate, a second plate and a spacer, wherein:

[0200] i. The panels are movable relative to each other into different configurations, including an open configuration and a closed configuration;

[0201] ii. each plate has a sample contact area on its respective sample surface for contacting a sample, wherein the sample contains the microorganisms to be analyzed, and

[0202] iii. one or both of the plates contain spacers fixed to the corresponding sample contact areas, the spacers have a predetermined height that is uniform, and at least one of the spacers is within the sample contact area,

[0203] wherein in an open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by a spacer, and the sample is deposited on one or both of the plates;

[0204] Wherein in the closed configuration, the closed configuration is configured after the sample is deposited in the open configuration: at least a portion of the sample is compressed by two plates into a layer of very uniform thickness, and the uniform thickness of the layer is defined by the sample contacting surfaces of the plates and adjusted by the plates and spacers.

[0205] B0. A system for examining the effect of a drug on a microorganism, comprising:

[0206] (a) the device described in any one of the embodiments;

[0207] (b) an imager comprising a camera and a light source for imaging microorganisms in a layer of uniform thickness; and

[0208] (c) A processor comprising electronics, a signal processor, and hardware and software for receiving and processing images and identifying and analyzing microorganisms in the images.

[0209] B1. A system for examining the effect of a drug on a microorganism, comprising:

[0210] (a) the device described in any one of the embodiments;

[0211] (b) a mobile communication device comprising:

[0212] i. an imager comprising one or more cameras for imaging microorganisms in a sample;

[0213] ii. a processor comprising electronics, a signal processor, and hardware and software for receiving and / or processing images of microorganisms and for remote communication; and

[0214] (c) a light source from the mobile communication device or an external source, wherein the light source is configured to provide illumination to the sample for imaging with the camera.

[0215] C1. A method for examining the effect of a drug on a microorganism, comprising the following steps:

[0216] (a) obtaining a sample, the sample containing the microorganism to be analyzed and mixing it with a test reagent;

[0217] (b) obtaining a first plate and a second plate movable relative to each other into different configurations, the different configurations comprising an open configuration and a closed configuration, wherein:

[0218] i. Each plate has a sample contact area on its respective surface for contacting the sample, and

[0219] ii. one or both of the plates contain a spacer fixed to the corresponding sample contact surface,

[0220] wherein the spacers have a predetermined substantially uniform height and at least one of the spacers is within the sample contacting region;

[0221] (c) depositing a sample on one or both of the plates while the plates are in an open configuration,

[0222] wherein in the open configuration, the two plates are partially or completely separated and the spacing between the plates is not adjusted by spacers;

[0223] (d) after (c), bringing the two plates together and pressing the plates into a closed configuration, wherein in the closed configuration: at least a portion of the sample is compressed by the two plates into a layer of very uniform thickness, the uniform thickness of the layer being defined by the sample surfaces of the two plates and adjusted by the spacers and the plates;

[0224] (e) When the plate is in the closed configuration, monitoring the microorganisms in the uniform thickness layer and determining the effect of the test agent on the microorganisms.

[0225] CC1. A method for examining the effect of a drug on a microorganism, comprising the following steps:

[0226] (a) obtaining a sample, the sample containing a microorganism to be analyzed;

[0227] (b) obtaining a first plate and a second plate movable relative to each other into different configurations, the different configurations comprising an open configuration and a closed configuration, wherein:

[0228] i. each plate has a sample contact area on its respective surface for contacting the sample,

[0229] ii. one or both of the plates contain a spacer fixed to the corresponding sample contact surface, and

[0230] iii. at least one of the plates contains a test reagent on a corresponding sample contact area, the test reagent dissolving and diffusing in the sample upon contact with the sample,

[0231] wherein the spacers have a predetermined substantially uniform height and at least one of the spacers is within the sample contacting region;

[0232] (c) depositing a sample on one or both of the plates while the plates are in an open configuration,

[0233] wherein in the open configuration, the two plates are partially or completely separated and the spacing between the plates is not adjusted by spacers;

[0234] (d) after (c), bringing the two plates together and pressing the plates into a closed configuration, wherein in the closed configuration: at least a portion of the sample is compressed by the two plates into a layer of very uniform thickness, the uniform thickness of the layer being defined by the sample surfaces of the two plates and adjusted by the spacers and the plates;

[0235] (e) When the plate is in the closed configuration, monitoring the microorganisms in the uniform thickness layer and determining the effect of the test agent on the microorganisms.

[0236] A2. An apparatus, system or method as described in any preceding embodiment, wherein one or both of the plates contain one or more test reagents on the corresponding sample contact areas, and the test reagents dissolve and diffuse in the sample when contacting the sample.

[0237] A3. An apparatus, system or method as described in any of the preceding embodiments, wherein one or both of the plates comprise a control zone and an experimental zone on the corresponding sample contact area, wherein the experimental zone comprises one or more test reagents that dissolve and diffuse in the sample when in contact with the sample, and wherein the control zone does not comprise such test reagents.

[0238] A4. An apparatus, system or method as described in any preceding embodiment, wherein the uniform height of the spacer is equal to or less than the average size of the microorganisms.

[0239] A5. An apparatus, system or method as described in any preceding embodiment, wherein the uniform height of the spacer is in the range of 75% to 125% of the average size of the microorganisms.

[0240] A6. An apparatus, system or method as described in any preceding embodiment, wherein at least one of the plates is transparent.

[0241] A7. An apparatus, system or method as described in any preceding embodiment, wherein one or both of the plates contain a dye on the corresponding sample contact area, and the dye dissolves in the sample and stains the microorganisms when it contacts the sample.

[0242] A8. An apparatus, system or method as described in Example A4, wherein the dye is fluorescently labeled.

[0243] A9. An apparatus, system or method as described in any of the preceding embodiments, wherein one or both of the plates contain a reactive dye on the corresponding sample contact area, the reactive dye being selected from the group consisting of: propidium iodide, 7-AAD, trypan blue, calcein violet AM, calcein AM, fixable reactive dyes, SYTO9 and other nucleic acid dyes, resazurin and colored formazan (MTT / XTT) and other mitochondrial dyes, and any combination thereof.

[0244] A10. The device, system or method of any preceding embodiment, wherein the sample comprises an agent that promotes the survival and / or division of microorganisms.

[0245] A11. An apparatus, system or method as described in any preceding embodiment, wherein one or both of the plates contain reagents that promote microbial survival and / or division on the corresponding sample contact areas.

[0246] B2. The system of any preceding embodiment, further comprising:

[0247] (d) A housing configured to hold a sample and to be mounted to a mobile communication device.

[0248] B3. The system of any preceding embodiment, wherein the imager is configured to obtain a series of images of the microorganism at discrete time points.

[0249] B4. A system as described in embodiment B3, wherein the processor is configured to receive and analyze the series of images and identify and count microorganisms in the first area of ​​each image.

[0250] B5. The system of any preceding embodiment, wherein the imager is configured to obtain a video comprising a series of images of the microorganism over a first period of time.

[0251] B6. A system as described in embodiment B5, wherein the processor is configured to receive and analyze the video, identify and track at least a portion of the microorganisms in the video, and evaluate changes in the tracked microorganisms over at least a portion of the first time period.

[0252] B7. A system as described in any of the preceding embodiments, wherein the housing includes an optical device for facilitating imaging and / or signal processing of a sample by a mobile communication device, and a base configured to hold the optical device on the mobile communication device.

[0253] B8. The system of any preceding embodiment, wherein the mobile communication device is configured to transmit the test results to medical personnel, a medical institution, or an insurance company.

[0254] B9. A system as described in any of the preceding embodiments, wherein the mobile communication device is further configured to communicate information about the subject with medical personnel, medical institutions, or insurance companies.

[0255] B10. The system of any preceding embodiment, wherein the mobile communication device is configured to receive a prescription, diagnosis, or advice from a medical professional.

[0256] B11. The system of any preceding embodiment, wherein the mobile communication device communicates with the remote location via WiFi or a cellular network.

[0257] B12. A system as described in any preceding embodiment, wherein the mobile communication device is a mobile phone.

[0258] B13. The system of any preceding embodiment, further comprising a temperature control element that maintains the sample deposited between the two plates at a temperature that promotes the survival and / or division of microorganisms.

[0259] C2. A method as described in any of the preceding embodiments, wherein the monitoring and de-determining step (e) is performed by a mobile communication device, the mobile communication device comprising:

[0260] i. an imager comprising one or more cameras for imaging microorganisms in a sample; and

[0261] ii. a processor comprising electronics, a signal processor, and hardware and software for receiving and / or processing images of microorganisms and for remote communication; and

[0262] A light source from a mobile communication device or an external source, wherein the light source is configured to provide illumination to the sample for imaging with the camera.

[0263] C3. The method of any preceding embodiment, wherein the monitoring and determining step (e) comprises:

[0264] i. Obtaining a series of images of platelets in a uniform thickness layer at discrete time points;

[0265] ii. identifying and counting microorganisms in at least one region of each image; and

[0266] iii. Assessing changes in the microorganisms between at least some discrete time points.

[0267] C4. The method of any preceding embodiment, wherein the monitoring and determining step (e) comprises:

[0268] i. obtaining a video comprising a series of images of a plate in a layer of uniform thickness over a first period of time;

[0269] ii. identifying and tracking at least a portion of one or more microorganisms in the video; and

[0270] iii. evaluating changes in the tracked microorganisms during at least a portion of the first time period.

[0271] C5. The method of any of preceding embodiments C3 or C4, wherein the identifying step comprises comprising microorganisms stained with a dye, and wherein the dye comprises a cell viability dye that indicates whether the stained cells are alive or dead.

[0272] C6. A method as described in Example C5, wherein the cell viability dye is selected from: propidium iodide, 7-AAD, trypan blue, calcein violet AM, calcein AM, fixable viability dyes, SYTO9 and other nucleic acid dyes, resazurin and colored formazan (MTT / XTT) and other mitochondrial dyes, and any combination thereof.

[0273] C7. The system of any preceding embodiment, further comprising:

[0274] When the plates are in the closed configuration, the sample is maintained at a temperature that is favorable for the survival and / or division of the microorganisms.

[0275] E1. A device, system or method as described in any preceding embodiment, wherein the spacer has:

[0276] i. having a substantially uniform cross-section and a flat top surface of the column shape;

[0277] ii. A width-to-height ratio equal to or greater than 1;

[0278] iii. a fill factor equal to or greater than 1%; and

[0279] iv. The product of the filling factor and the Young's modulus of the spacer is 2 MPa or more.

[0280] Where fill factor is the ratio of the spacer contact area to the total board area.

[0281] E2. An apparatus, system or method as described in any preceding embodiment, wherein the average value of the uniform thickness of the layer is substantially the same as the uniform height of the spacers, with a variation of less than 10%.

[0282] E3. An apparatus, system or method as described in any preceding embodiment, wherein the uniform thickness layer varies by less than 30 nm.

[0283] E4. An apparatus, system or method as described in any preceding embodiment, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 5%.

[0284] E5. A device, system or method as described in any preceding embodiment, wherein the spacer has a column with a cross-sectional shape selected from the following: circular, polygonal, annular, square, rectangular, oval, elliptical or any combination thereof.

[0285] E6. A device, system or method as described in any preceding embodiment, wherein the spacer has:

[0286] i. having a substantially uniform cross-section and a flat top surface of the column shape;

[0287] ii. A width-to-height ratio equal to or greater than 1;

[0288] iii. a predetermined constant spacer spacing in the range of 10m to 200Dm;

[0289] iv. a fill factor equal to or greater than 1%; and

[0290] v. The product of the filling factor and the Young's modulus of the spacer is 2 MPa or more.

[0291] Where fill factor is the ratio of the spacer contact area to the total board area.

[0292] E7. An apparatus, system or method as described in any of the preceding embodiments, wherein pressing the plate into a closed structure is performed in parallel or sequentially, parallel pressing applies external force on the intended area simultaneously, and sequential pressing applies external force on a portion of the intended area and gradually moves to other areas.

[0293] E8. An apparatus, system or method as described in any preceding embodiment, wherein the uniform thickness sample layer has a thickness uniformity of up to + / - 5%.

[0294] Related Documents

[0295] 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 the 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 document, but also the documents cited, incorporated or claiming priority here.

[0296] (1) definition

[0297] The terms used to describe the devices, systems, and methods disclosed herein are defined in this application or in PCT Application Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0298] (2) Spacers and uniformity

[0299] The devices, systems and methods disclosed herein may include or use a QMAX card for sample detection, analysis and quantification. In some embodiments, the QMAX card includes a spacer that helps to make at least a portion of the sample a very uniform layer. The structure, materials, functions, variations and sizes of the spacer, and the uniformity of the spacer and sample layer are disclosed 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, and U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0300] (3) Hinges, notches, grooves and slides

[0301] The devices, systems, and methods disclosed herein may include or use a QMAX card for sample detection, analysis, and quantification. In some embodiments, the QMAX card includes hinges, notches, grooves, and sliders that help facilitate the operation of the QMAX card and the measurement of samples. The structures, materials, functions, variations, and dimensions of the hinges, notches, grooves, and sliders are disclosed herein, or listed, described, and summarized in PCT Applications (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and in U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, all of which are incorporated herein by reference in their entirety.

[0302] (4) Card, Slider and Smartphone Inspection Systems

[0303] The devices, systems, and methods disclosed herein may include or use a QMAX card for sample detection, analysis, and quantification. In some embodiments, the QMAX card is used with a slider of the card that allows a smart card detection system to read. The structure, materials, functions, variations, dimensions, and connections of the QMAX card, slider, and smart phone detection system are disclosed herein or listed, described, and summarized in PCT Applications (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0304] (5) Detection Methods

[0305] 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 applications (designated U.S.) No. PCT / US2016 / 045437 and No. PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and U.S. Provisional Application No. 62 / 456065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0306] (6) Label

[0307] The devices, systems and methods disclosed herein may use various types of tags. Tags are disclosed herein or listed, described and summarized in PCT Applications (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0308] (7) Biomarkers

[0309] The devices, systems and methods disclosed herein can use various types of biomarkers. Biomarkers 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, and U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0310] (8) cloud

[0311] The devices, systems and methods disclosed herein may use cloud technology for data transmission, storage and / or analysis. Relevant cloud technologies are disclosed or listed, described and summarized in PCT applications (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, and U.S. Provisional Application No. 62 / 456,065, filed on February 7, 2017, the entire contents of which are incorporated herein by reference.

[0312] (9) Applications (fields and samples)

[0313] Device, system and method disclosed herein can be used for various applications (field and sample).Described application is disclosed herein or lists, describes and is summarized in PCT application (designated U.S.) No. PCT / US2016 / 045437 and No. PCT / US0216 / 051775, respectively submitted on August 10, 2016 and September 14, 2016, and in U.S. Provisional Application No. 62 / 456065, submitted on February 7, 2017, the entire contents of these applications are incorporated herein by reference.

[0314] Other Notes

[0315] Other embodiments according to the inventive subject matter of the present disclosure are described in the following enumerated paragraphs.

[0316] 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.

[0317] As used herein, the term "analyte" refers to molecules (e.g., proteins, peptides, DNA, RNA, nucleic acids or other molecules) with different shapes, cells, tissues, viruses and nanoparticles. It can also be referred to as any substance suitable for testing in the present invention.

[0318] 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.

[0319] 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.

[0320] 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.

[0321] 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.

[0322] 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.

[0323] 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.

[0324] 1. sample

[0325] The devices, apparatus, systems and methods disclosed herein can be used for samples, such as but not limited to diagnostic samples, clinical samples, environmental samples and food samples. The types of samples include but are not limited to the samples listed, described and / or summarized in PCT applications (designating the United States) Nos. PCT / US2016 / 045437 and PCT / US0216 / 051775, filed on August 10, 2016 and September 14, 2016, respectively, the entire contents of which are incorporated herein by reference.

[0326] For example, in some embodiments, the devices, apparatus, systems and methods disclosed herein are used for samples comprising cells, tissues, body fluids and / or mixtures thereof. In some embodiments, the sample comprises a human body fluid. In some embodiments, the sample comprises at least one of the following: cells, tissues, body fluids, feces, amniotic fluid, aqueous humor, vitreous humor, blood, whole blood, fractionated blood, plasma, serum, breast milk, cerebrospinal fluid, cerumen, chyle, chyme, endolymph, perilymph, excrement, gastric acid, gastric juice, lymph, mucus, nasal drainage, sputum, pericardial fluid, peritoneal fluid, pleural fluid, pus, rheumatic fluid, saliva, sebum, semen, sputum, sweat, synovial fluid, tears, vomitus, urine and exhaled breath condensate.

[0327] In some embodiments, the devices, apparatuses, systems and methods disclosed herein are used for environmental samples obtained from any suitable source, such as, but not limited to, liquid samples from rivers, lakes, ponds, oceans, glaciers, icebergs, rainwater, snow, sewage, reservoirs, tap water, drinking water, etc.; solid samples from soil, compost, sand, rock, concrete, wood, brick, dirt, etc.; and gas samples from air, underwater heat dissipation, industrial exhaust, vehicle exhaust, etc. In certain embodiments, the environmental sample is a fresh sample obtained from a source; in certain embodiments, the environmental sample is processed. For example, before applying the subject apparatus, equipment, system and method, the sample in non-liquid form is converted into a liquid form.

[0328] In some embodiments, the devices, apparatuses, systems and methods disclosed herein are used for food samples that are suitable or may become suitable for animal consumption, such as human consumption. In some embodiments, food samples may include raw materials, cooked or processed foods, foods of plant and animal origin, pre-processed foods, and partially or completely processed foods, etc. In certain embodiments, samples in non-liquid form are converted to liquid form prior to application of the subject devices, apparatuses, systems and methods.

[0329] The devices, apparatus, systems and methods of the present invention can be used to analyze samples of any volume. Examples of volumes include, but are not limited to, about 10 mL or less, 5 mL or less, 3 mL or less, 1 microliter (μL, also referred to as "uL") or less, 500 μL or less, 300 μL or less, 250 μL or less, 200 μL or less, 170 μL or less, 150 μL or less, 125 μL or less, 100 μL or less, 75 μL or less, 50 μL or less, 25 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, 1 pL or less, or in a range between any two of said values.

[0330] In some embodiments, the volume of the sample includes but is not limited to about 100 μL or less, 75 μL or less, 50 μL or less, 25 μL or less, 20 μL or less, 15 μL or less, 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, 1 pL or less, or in a range between any two of said values. In some embodiments, the volume of the sample includes but is not limited to about 10 μL or less, 5 μL or less, 3 μL or less, 1 μL or less, 0.5 μL or less, 0.1 μL or less, 0.05 μL or less, 0.001 μL or less, 0.0005 μL or less, 0.0001 μL or less, 10 pL or less, 1 pL or less, or in a range between any two of said values.

[0331] In some embodiments, the amount of sample is about one drop of liquid. In some embodiments, the amount of sample is the amount collected from a punctured finger or fingertip. In some embodiments, the amount of sample is the amount collected from a microneedle, micropipette, or intravenous aspiration.

[0332] In certain embodiments, the sample holder is configured to hold a fluid sample. In certain embodiments, the sample holder is configured to compress at least a portion of the fluid sample into a thin layer. In certain embodiments, the sample holder includes a structure configured to heat and / or cool the sample. In certain embodiments, a heating source provides electromagnetic waves, which can be absorbed by certain structures in the sample holder to change the temperature of the sample. In certain embodiments, a signal sensor is configured to detect and / or measure a signal from a sample. In certain embodiments, a signal sensor is configured to detect and / or measure an analyte in a sample. In certain embodiments, a radiator is configured to absorb heat from a sample holder and / or a heating source. In certain embodiments, a radiator includes a chamber that at least partially surrounds the sample holder.

[0333] 2. application

[0334] The devices, apparatus, systems and methods disclosed herein can be used for various types of biological / chemical sampling, sensing, assays and applications, including PCT application (designating the United States) No. PCT / US2016 / 045437, which was filed on August 10, 2016 and is incorporated herein by reference in its entirety.

[0335] In some embodiments, the devices, apparatuses, systems and methods disclosed herein are used in a variety of different applications in a variety of fields, where it is necessary to determine the presence or absence, quantify and / or amplify one or more analytes in a sample. For example, in certain embodiments, the subject devices, apparatuses, systems and methods are used to detect proteins, peptides, nucleic acids, synthetic compounds, inorganic compounds, organic compounds, bacteria, viruses, cells, tissues, nanoparticles and other molecules, compounds, mixtures and substances thereof. The various fields in which the subject devices, apparatuses, systems and methods can be used include, but are not limited to: diagnosis, management and / or prevention of human diseases and conditions, diagnosis, management and / or prevention of animal diseases and conditions, diagnosis, management and / or prevention of plant diseases and conditions, agricultural uses, veterinary uses, food testing, environmental testing and purification, drug testing and prevention, etc.

[0336] Applications of the present invention include, but are not limited to: (a) detection, purification, quantification and / or amplification of compounds or biomolecules associated with certain diseases or certain stages of diseases, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancers, mental disorders, neuropsychiatric disorders and organ diseases, such as lung diseases, kidney diseases, (b) detection, purification, quantification and / or amplification of cells and / or microorganisms from the environment, such as viruses, fungi and bacteria, the environment is such as water, soil or biological samples, such as tissues, body fluids, (c) detection, quantification of compounds or biological samples that pose a threat to food safety, human health or national security, such as toxic waste, anthrax, (d) detection and quantification of vital parameters in medical or physiological monitors, such as glucose, blood oxygen level, total blood cell count, (e) detection and quantification of specific DNA or RNA from biological samples (such as cells, viruses, body fluids), (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genomic analysis, or (g) detection and quantification of reaction products, such as during the synthesis or purification of drugs.

[0337] In some embodiments, the subject device, apparatus, system and method are used to detect nucleic acids, proteins or other molecules or compounds in a sample. In certain embodiments, the device, apparatus, system and method are used to quickly, clinically detect and / or quantify one or more, two or more, or three or more disease biomarkers in a biological sample, for example, for diagnosing, preventing and / or controlling a disease condition in a subject. In certain embodiments, the device, apparatus, system and method are used to detect and / or quantify one or more, two or more, or three or more environmental markers in an environmental sample, such as a sample obtained from a river, ocean, lake, rain, snow, sewage, sewage treatment runoff, agricultural runoff, industrial runoff, tap water or drinking water. In certain embodiments, the device, apparatus, system and method are used to detect and / or quantify one or more, two or more, or three or more food markers from a food sample, and the food sample is obtained from tap water, drinking water, prepared food, processed food or unprocessed food.

[0338] In some embodiments, the subject device is part of a microfluidic device. In some embodiments, the subject device, apparatus, system and method are used to detect fluorescent or luminescent signals. In some embodiments, the subject device, apparatus, system and method include or are used with a communication device, such as but not limited to a mobile phone, a tablet computer and a portable computer. In some embodiments, the subject device, apparatus, system and method include or are used with an identifier, such as but not limited to an optical barcode, a radio frequency ID tag or a combination thereof.

[0339] In some embodiments, the sample is a diagnostic sample obtained from a subject, the analyte is a biomarker, and the amount of the analyte in the sample measured can be diagnostic of a disease or condition. In some embodiments, the subject device, system, and method further include receiving or providing a report to the subject indicating the measured amount of the biomarker in an individual who does not have or is at low risk of having a disease or condition and a range of measured values ​​for the biomarker, wherein the measured amount of the biomarker relative to the range of measured values ​​is used to diagnose the disease or condition.

[0340] In some embodiments, the sample is an environmental sample, and wherein the analyte is an environmental marker. In some embodiments, the subject device, system and method include receiving or providing a report indicating the safety or hazard of the subject exposed to the environment from which the sample is obtained. In some embodiments, the subject device, system and method include sending data containing the amount of the measured environmental marker to a remote location and receiving a report indicating the safety or hazard of the subject exposed to the environment from which the sample is obtained.

[0341] In some embodiments, the sample is a food sample, wherein the analyte is a food marker, and wherein the amount of the food marker in the sample is related to the safety of consuming the food. In some embodiments, the subject device, system, and method include receiving or providing a report indicating the safety or hazard of the subject consuming the food from which the sample was obtained. In some embodiments, the subject device, system, and method include sending data containing a measured amount of a food marker to a remote location and receiving a report indicating the safety or hazard of the subject consuming the food from which the sample was obtained.

[0342] 3. Analytes, Biomarkers, and Diseases

[0343] The devices, apparatus, systems and methods disclosed herein can be used for the detection, purification and / or quantification of various analytes. In some embodiments, the analytes are biomarkers associated with various diseases. In some embodiments, the analytes and / or biomarkers indicate the presence, severity and / or stage of a disease. Analytes, biomarkers and / or diseases that can be detected and / or measured using the devices, apparatus, systems and / or methods of the present invention include those listed, described and / or summarized in PCT Application No. PCT / US2016 / 045437 filed on August 10, 2016 (designating the United States), No. PCT / US2016 / 054025 filed on September 27, 2016, and U.S. Provisional Application No. 62 / 234,538 filed on September 29, 2015, No. 62 / 233,885 filed on September 28, 2015, No. 62 / 293,188 filed on February 9, 2016, and No. 62 / 305,123 filed on March 8, 2016, which applications are hereby incorporated by reference in their entirety. For example, the devices, apparatus, systems and methods disclosed herein can be used for (a) detection, purification and quantification of compounds or biomolecules associated with certain disease stages, such as infectious and parasitic diseases, injuries, cardiovascular diseases, cancers, mental disorders, neuropsychiatric disorders and organic diseases, such as lung disease, kidney disease, (b) detection, purification and quantification of microorganisms such as viruses, fungi and bacteria from the environment, such as water, soil or biological samples, such as tissues and body fluids, (c) detection and quantification of chemical compounds or biological samples that pose a threat to food safety or national security, such as toxic waste, anthrax, (d) quantification of vital parameters in medical or physiological monitors, such as glucose, blood oxygen levels, total blood cell counts, (e) detection and quantification of specific DNA or RNA from biological samples, such as cells, viruses, body fluids, (f) sequencing and comparison of genetic sequences of DNA in chromosomes and mitochondria for genomic analysis, or (g) detection of reaction products, such as during drug synthesis or purification.

[0344] In some embodiments, the analyte can be a biomarker, an environmental marker, or a food marker. In some cases, the sample is a liquid sample and can be a diagnostic sample (e.g., saliva, serum, blood, sputum, urine, sweat, tears, semen, or mucus); an environmental sample obtained from a river, ocean, lake, rainwater, snow, sewage, sewage runoff, agricultural runoff, industrial runoff, tap water, or drinking water; or a food sample obtained from tap water, drinking water, prepared food, processed food, or unprocessed food.

[0345] In any embodiment, the sample can be a diagnostic sample obtained from a subject, the analyte can be a biomarker, and the amount of the analyte in the sample measured can be diagnostic of a disease or condition.

[0346] In any embodiments, the devices, apparatus, systems and methods of the present invention may further include diagnosing a subject based on information including measured amounts of biomarkers in a sample. In some cases, the diagnosing step includes sending data including the measured amounts of biomarkers to a remote location and receiving a diagnosis based on the information including the measurements from the remote location.

[0347] In any embodiment, the biomarker can be selected from Table B1, 2, 3 or 7 disclosed in U.S. Provisional Application Nos. 62 / 234,538, 62 / 293,188 and / or 62 / 305,123 and / or PCT Application No. PCT / US2016 / 054,025, the entire contents of which are incorporated herein for all purposes. In some cases, the biomarker is a protein selected from Table B1, 2 or 3. In some cases, the biomarker is a nucleic acid selected from Table B2, 3 or 7. In some cases, the biomarker is a biomarker derived from an infectious agent selected from Table B2. In some cases, the biomarker is a microRNA (miRNA) selected from Table B7.

[0348] In any embodiment, applying step b) can include isolating miRNA from the sample to produce an isolated miRNA sample, and applying the isolated miRNA sample to a disk-coupled dot-rod antenna (QMAX device) array.

[0349] In any embodiment, the QMAX device may contain a plurality of capture agents that each bind a biomarker selected from Tables B1, B2, B3, and / or B7, wherein the reading step d) comprises obtaining a measurement of the amount of the plurality of biomarkers in the sample, and wherein the amount of the plurality of biomarkers in the sample is diagnostic of a disease or condition.

[0350] In any embodiment, the capture agent can be an antibody epitope and the biomarker can be an antibody that binds to the antibody epitope. In some embodiments, the antibody epitope comprises a biomolecule or a fragment thereof selected from Table B4, B5 or B6. In some embodiments, the antibody epitope comprises an allergen or a fragment thereof selected from Table B5. In some embodiments, the antibody epitope comprises a biomolecule or a fragment thereof derived from an infectious agent selected from Table B6.

[0351] In any embodiment, the QMAX device can contain a plurality of antibody epitopes selected from Tables B4, B5 and / or B6, wherein the reading step d) comprises obtaining a measurement of the amount of the plurality of epitope binding antibodies in the sample, and wherein the amount of the plurality of epitope binding antibodies in the sample is diagnostic of a disease or condition.

[0352] In any embodiment, the sample can be an environmental sample, and the analyte can be an environmental marker. In some embodiments, the environmental marker is selected from Table B8 in U.S. Provisional Application No. 62 / 234,538 and / or PCT Application No. PCT / US2016 / 054025.

[0353] In any embodiment, the method may include receiving or providing a report indicating the safety or harmfulness to the subject of exposure to the environment from which the sample was obtained.

[0354] In any embodiment, the method can include transmitting data including the measured amount of the environmental marker to a remote location and receiving a report indicating the safety or harmfulness to a subject exposed to the environment from which the sample was obtained.

[0355] In any embodiment, the QMAX device array can include a plurality of capture agents, each capture agent binding an environmental marker selected from Table B8, and wherein the reading step d) can include obtaining a measurement of the amount of the plurality of environmental markers in the sample.

[0356] In any embodiment, the sample can be a food sample, wherein the analyte can be a food marker, and wherein the amount of the food marker in the sample can be related to the safety of the edible food. In some embodiments, the food marker is selected from Table B9.

[0357] In any embodiment, the method can include receiving or providing a report indicating the safety or harmfulness of the food from which the sample was obtained by the subject.

[0358] In any embodiment, the method can include sending data containing the measured amounts of food markers to a remote location and receiving a report indicating the safety or harmfulness of the subject consuming the food from which the sample was obtained.

[0359] In any embodiments, the devices, apparatus, systems and methods disclosed herein may include a plurality of capture agents that each bind a food marker selected from Table B9, Table B9 from U.S. Provisional Application No. 62 / 234,538 and PCT Application No. PCT / US2016 / 054025, wherein the obtaining may include obtaining measurements of the amounts of the plurality of food markers in the sample, and wherein the amounts of the plurality of food markers in the sample may be correlated to the safety of the food for consumption.

[0360] Also provided herein are kits for practicing the devices, systems, and methods of the invention.

[0361] The amount of sample can be about one drop of sample. The amount of sample can be the amount collected from a finger prick or. The amount of sample can be the amount collected from a microneedle or intravenous aspiration.

[0362] The sample may be used without further processing after being obtained from the source, or may be processed, for example, to enrich for an analyte of interest, remove large particulate matter, dissolve or resuspend a solid sample, etc.

[0363] Any suitable method of applying the sample to the QMAX device may be used. Suitable methods may include using a pipette, dropper, syringe, etc. In certain embodiments, when the QMAX device is in a holder in the form of a dipstick, as described below, the sample may be applied to the QMAX device by dipping the sample receiving area of ​​the dipstick into the sample.

[0364] Samples may be collected one or more times. Samples collected over time may be aggregated and / or processed individually (as described herein, by application to a QMAX device and obtaining measurements of the amount of analyte in the sample). In some cases, measurements obtained over time may be aggregated and used for longitudinal analysis over time to facilitate screening, diagnosis, treatment, and / or disease prevention.

[0365] Washing the QMAX device to remove unbound sample components can be performed in any convenient manner, as described above. In certain embodiments, the surface of the QMAX device is washed with a binding buffer to remove unbound sample components.

[0366] Detectable labeling of the analyte can be performed by any convenient method. The analyte can be labeled directly or indirectly. In direct labeling, the analyte in the sample is labeled before the sample is applied to the QMAX device. In indirect labeling, the unlabeled analyte in the sample is labeled after the sample is applied to the QMAX device to capture the unlabeled analyte, as described below.

[0367] 4. Label

[0368] The devices, apparatus, systems, and methods disclosed herein can be used with various types of tags, including tags disclosed, described, and / or summarized in PCT Application No. (designating the United States) PCT / US2016 / 045437, which was filed on August 10, 2016 and is incorporated herein by reference in its entirety.

[0369] In some embodiments, the label is optically detectable, such as, but not limited to, a fluorescent label. In some embodiments, the label includes, but is not limited to, IRDye800CW, Alexa 790, Dylight 800, fluorescein, fluorescein isothiocyanate, succinimidyl ester of carboxyfluorescein, succinimidyl ester of fluorescein, 5-isomer of fluorescein dichlorotriazine, caged carboxyfluorescein-alanine-carboxamide, Oregon Green 488, Oregon Green 514; fluorescein yellow, acridine orange, rhodamine, tetramethylrhodamine, Texas Red, propidium iodide, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide), tetrabromorhodamine 123, rhodamine 6G, TMRM (tetramethylrhodamine methyl ester), TMRE (tetramethylrhodamine ethyl ester), tetramethylrhodamine, rhodamine B and 4-dimethylaminotetramethylrhodamine. Ming, green fluorescent protein, blue-shifted green fluorescent protein, blue-green-shifted green fluorescent protein, red-shifted green fluorescent protein, yellow-shifted green fluorescent protein, 4-acetamido-4'-isothiocyanate stilbene-2,2'-disulfonic acid; acridine and derivatives, for example, acridine, acridine isothiocyanate; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS); 4-amino-N-[3-vinylsulfonyl)phenyl]naphthalenamide-3,5-disulfonate; N-(4-anilino-1-naphthyl)maleimide; anthranilamide; 4,4-difluoro-5-(2-thienyl)-4-boron-3a,4adiaza-5-indacene-3-propionic acid BODIPY; grade Blue; Brilliant Yellow; Coumarin and derivatives: Coumarin, 7-amino-4-methylcoumarin (AMC, Coumarin 120), 7-amino-4-methylcoumarin (AMC, Coumarin 151); Cyanine dyes; cyanosine; 4',6-diamino-2-phenylindole (DAPI); 5',5"-dibromopyrogallolsulfonaphthalene (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; Diethylenetriamine pentaacetate; 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid; 4,4'-diisothiocyanato-2,2'-yldisulfonic acid disodium salt; 5-(dimethylamino)naphthalene-1-sulfonyl Chloride (DNS, dansyl chloride); 4-dimethylaminophenylazophenyl-4'-isothiocyanate (DABITC); Eosin and derivatives: Eosin, Eosin isothiocyanate, Erythrophyca and derivatives: Erythrophyca B, Erythrophyca, isothiocyanate; Ethidium; Fluorescein and its derivatives: 5-carboxyfluorescein (FAM), 5-(4,6-dichlorotriazin-2-yl)amino-fluorescein (DTAF), 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein (JOE), Fluorescein, Fluorescein isothiocyanate, QFITC, (XRITC); Fluoresceinamine; IR144; IR1446; Malachite green isothiocyanate; 4-methylumbelliferyl o-cresolphthalein;Nitrotyrosine; para-carmine; phenol red; B-phycoerythrin; o-phthalaldehyde; pyrene and its derivatives: pyrene, pyrenebutyric acid, succinimidyl 1-pyrene; butyrate quantum dots; Reactive Red 4 (Cibacron; TM Brilliant Red 3B-A) Rhodamine and derivatives: 6-carboxy-X-rhodamine (ROX), 6-carboxyrhodamine (R6G), lissamine rhodamine B sulfonyl chloride rhodamine (Rhod), rhodamine B, rhodamine 123, rhodamine X isothiocyanate, sulforhodamine B, sulforhodamine 101, sulfonyl chloride derivatives of 5-sulforhodamine (Texas Red); N,N,N',N'-tetramethyl-6-carboxyrhodamine TAMRA; tetramethylrhodamine; tetramethylrhodamine isothiocyanate (TRITC); riboflavin; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS), 4-(4'-dimethylaminophenylazo)benzoic acid (DABCYL), rhodamine; CAL fluorescent orange 560; terbium chelate derivatives; Cy3; Cy5; Cy5.5; Cy7; IRD700; IRD 800; La Jolla Blue; phthalocyanines; and naphthalocyanines, coumarins and related dyes, xanthene dyes such as rhodols, resorufins, bimanes, acridines, isoindoles, dansyl dyes, aminophthalhydrazides (e.g., luminol), and isoluminol derivatives, aminophthalimides, aminonaphthalimides, aminobenzofurans, aminoquinosides, dicyanohydroquinones, fluorescent europium and terbium complexes; combinations thereof, and the like. Suitable fluorescent and chromogenic proteins include, but are not limited to, green fluorescent protein (GFP), including, but not limited to, GFP derived from Aequoria victoria or a derivative thereof, e.g., a "humanized" derivative, such as enhanced GFP; GFP from another species, such as Renilla reniformis, Renilla mulleri, or Ptilosarcus guernyi; "humanized" recombinant GFP (hrGFP); any of a variety of fluorescent and colored proteins from coral species; any combination thereof; and the like.

[0370] 5. QMAX Device

[0371] The devices, apparatus, systems and methods disclosed herein may include or use a QMAX device ((Q: quantification; M: amplification; A: addition of reagents; X: acceleration; also referred to in some embodiments as a Q card or a compression regulated open flow (CROF) device), including the QMAX devices listed, described and / or summarized in PCT Application (designating the United States) No. PCT / US2016 / 045437 filed on August 10, 2016 and U.S. Provisional Application No. 62,431,639 filed on December 9, 2016 and No. 62 / 456,287 filed on February 8, 2017, the entire contents of which are incorporated herein by reference.

[0372] As used herein, 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 is described in Provisional Application Serial No. 62 / 456,065 filed on February 7, 2017, the entire contents of which are incorporated herein for all purposes.

[0373] The term "Compression Open Flow (COF)" refers to a method of changing the shape of a flowable sample deposited on a plate by (i) placing another plate on top of at least a portion of the sample, and (ii) then compressing the sample between two plates by pushing the two plates toward each other; wherein the compression reduces the thickness of at least a portion of the sample and causes the sample to flow into the open space between the plates. The term "Compression Regulated Open Flow" or "CROF" (or "Self-calibrated Compression Open Flow" or "SCOF" or "SCCOF") (also known as QMAX) refers to a specific type of COF in which the final thickness of part or all of the sample after compression is "regulated" by a spacer, which is placed between the two plates. Here, the CROF device can be used interchangeably with the QMAX card.

[0374] In the QMAX process, the term "open configuration" of two plates refers to a structure in which the two plates are either partially or completely separated and the spacing between the plates is not adjusted by spacers.

[0375] The term "closed configuration" of two plates in QMAX processing refers to a configuration in which the plates face each other, a spacer and a relevant volume of the sample are between the plates, the relevant spacing between the plates and therefore the thickness of the relevant volume of the sample are adjusted by the plates and the spacer, wherein the relevant volume is at least a portion of the entire volume of the sample.

[0376] In the QMAX process, the term "sample thickness is adjusted by the plate and spacer" means that, for given conditions of the plate, sample, spacer and plate compression method, the thickness of at least one port of the sample in the closed configuration of the plate can be predetermined according to the properties of the spacer and the plate.

[0377] The term "inner surface" or "sample surface" of the plate in the QMAX card refers to the surface of the plate that contacts the sample, while the other surface of the plate (not contacting the sample) is called the "outer surface".

[0378] Unless otherwise specified, the terms "height" or "thickness" of an object in the QMAX process refer to the dimension of the object in the direction perpendicular to the board surface. For example, the spacer height is the dimension of the spacer in the direction perpendicular to the board surface, and the spacer height and spacer thickness mean the same thing.

[0379] Unless otherwise specified, the term "area" of an object in the QMAX process refers to the area of ​​the object parallel to the plate surface. For example, a spacer area is a spacer area parallel to the plate surface.

[0380] The term QMAX card refers to a device that performs a QMAX (eg, CROF) process on a sample, and may or may not have a hinge connecting the two plates.

[0381] The terms "QMAX card with hinge" and "QMAX card" are interchangeable.

[0382] The terms "angle self-maintaining", "angle self-maintaining" or "rotation angle self-maintaining" refer to a property of a hinge that substantially maintains the angle between two panels after an external force moving the panels from an initial angle to the angle is removed from the panels.

[0383] When using the QMAX card, both plates need to be opened first for sample deposition. However, in some embodiments, the QMAX card from the package has two plates that are in contact with each other (e.g., closed position), and because one or both plates are very thin, it is a challenge to separate them. To facilitate the opening of the QMAX card, one or more opening notches are created at the edge or corner of the first plate or at these two locations, and in the closed position of the plates, a portion of the second plate is placed over the opening notches, so that in the notch of the first plate, the second plate can be lifted open without blocking the first plate.

[0384] In the QMAX assay platform, the QMAX card uses two plates to manipulate the shape of the sample into a thin layer (e.g., by compression). In certain embodiments, plate manipulation requires multiple changes in the relative position of the two plates (referred to as: plate configuration) by human hands or other external forces. The QMAX card needs to be designed to make manual manipulation easy and fast.

[0385] In the QMAX assay, one of the plate configurations is an open configuration, in which the two plates are fully or partially separated (the spacing between the plates is not controlled by a spacer) and the sample can be deposited. The other configuration is a closed configuration, in which at least a portion of the sample deposited in the open configuration is compressed by the two plates into a layer of very uniform thickness, the uniform thickness of the layer being defined by the inner surfaces of the plates and regulated by the plates and the spacer. In some embodiments, the average spacing between the two plates is greater than 300 μm.

[0386] In the operation of the QMAX assay, the operator needs to first put the two plates in an open configuration ready for sample deposition, then deposit the sample on one or both plates, and finally close the plates to a closed position. In certain embodiments, the two plates of the QMAX card are initially on top of each other and need to be separated to enter the open configuration for sample deposition. When one of the plates is a thin plastic film (175 μm thick PMA), this separation is difficult to perform by hand. The present invention is intended to provide devices and methods that make the operation of certain assays (such as QMAX card assays) easy and fast.

[0387] In some embodiments, the QMAX device includes a hinge that connects two or more panels together so that the panels can be opened and closed in a manner similar to a book. In some embodiments, the material of the hinge enables the hinge to self-maintain the angle between the panels after adjustment. In some embodiments, the hinge is configured to hold the QMAX card in a closed configuration so that the entire QMAX card can slide in and out of the card slot without causing accidental separation of the two panels. In some embodiments, the QMAX device includes one or more hinges capable of controlling the rotation of more than two panels.

[0388] In some embodiments, the hinge is made of a metal material selected from gold, silver, copper, aluminum, iron, tin, platinum, nickel, cobalt, alloys, or any combination thereof. In some embodiments, the hinge comprises a single layer made of a polymer material, such as, but not limited to, plastic. The polymer material is selected from the group consisting of: acrylate polymers, vinyl polymers, olefin polymers, cellulosic polymers, non-cellulosic polymers, polyester polymers, nylon, cyclic olefin copolymers (COC), poly(methyl methacrylate) (PMMB), polycarbonate (PC), cyclic olefin polymers (COP), liquid crystal polymers (LCP), polyamide (PB), polyethylene (PE), polyimide (PI), polypropylene (PP), polyphenylene ether (PPE), polystyrene (PS), polyoxymethylene (POM), polyetheretherketone (PEEK), polyethersulfone (PES), polyethylene phthalate (PET), polytetrafluoroethylene (PTFE), polyvinyl chloride (PVC), polyvinylidene fluoride (PVDF), polybutylene terephthalate (PBT), fluorinated ethylene propylene (FEP), perfluoroalkoxyalkane (PFB), polydimethylsiloxane (PDMS), rubber, or any combination thereof. In some embodiments, the polymer material is selected from polystyrene, PMMB, PC, COC, COP, other plastics, or any combination thereof.

[0389] In some embodiments, the QMAX device includes an opening mechanism, such as but not limited to notches on the edge of the board or strips attached to the board, making it easier for the user to manipulate the positioning of the board, such as but not limited to separating the boards by hand.

[0390] In some embodiments, the QMAX device comprises grooves on one or both plates. In certain embodiments, the grooves restrict the flow of samples on the plates.

[0391] 6. Spacers

[0392] The devices, apparatus, systems and methods disclosed herein may include or use a device (e.g., a QMAX device) that includes a spacer listed, described and / or summarized in PCT Application (designating the United States) No. PCT / US2016 / 045437 filed on August 10, 2016 and U.S. Provisional Application No. 62,431,639 filed on December 9, 2016 and No. 62 / 456,287 filed on February 8, 2017, the entire contents of which are incorporated herein by reference.

[0393] Essentially, unless otherwise specified, the term "spacer" or "stopper" refers to a mechanical object that, when placed between two plates, sets a limit on the minimum spacing between the two plates that can be reached when the two plates are compressed together. That is, during compression, the spacer will stop the relative motion of the two plates to prevent the plate spacing from becoming less than a preset (i.e., predetermined) value.

[0394] The terms "spacers have a predetermined height" and "spacers have a predetermined spacer spacing" mean that the values ​​of the spacer height and the spacer spacing, respectively, are known prior to the QMAX process. If the values ​​of the spacer height and the spacer spacing are not known prior to the QMAX process, the values ​​of the spacer height and the spacer spacing are not predetermined. For example, in the case of beads sprayed on the board as spacers, where the beads land at random locations on the board, the distance between the spacers is not predetermined. Another example of a non-predetermined inter-spacer distance is that the spacers move during the QMAX process.

[0395] In the QMAX process, the term "a spacer is fixed to its corresponding board" means that the spacer is connected to a position on the board and remains connected to that position during the QMAX process (i.e., the position of the spacer on the corresponding board does not change). An example of "a spacer is fixed to its corresponding board" is that the spacer is made integrally from one piece of material of the board and the position of the spacer relative to the board surface does not change during the QMAX process. An example of "a spacer is not fixed to its corresponding board" is that the spacer is bonded to the board by an adhesive, but during use of the board, during the QMAX process, the adhesive cannot hold the spacer in its original position on the board surface and the spacer moves away from its original position on the board surface.

[0396] 7. adapter

[0397] The devices, apparatus, systems and methods disclosed herein can be used with an adapter configured to accommodate the device and connect the device to a reader, such as but not limited to a smart phone. In some embodiments, the Q card is used with a slider that allows the card to be inserted into the adapter so that the card can be read by a smart phone detection system. The structure, materials, functions, variations, dimensions and connections of the Q card, slider and adapter are disclosed, listed, described and / or summarized in PCT Application Nos. PCT / US2016 / 045437 (designating the United States) filed on August 10, 2016 and PCT / US0216 / 051775 filed on September 14, 2016, U.S. Provisional Application Nos. 62 / 456,590 filed on February 8, 2017, 62 / 459,554 filed on February 15, 2017, and 62 / 460,075 filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0398] In some embodiments, the adapter includes a socket slot configured to accommodate the QMAX device when the device is in a closed configuration. In some embodiments, the QMAX device has a sample stored therein, and the adapter can be connected to a mobile device (e.g., a smart phone) so that the sample can be read by the mobile device. In some embodiments, the mobile device can detect and / or analyze signals from the sample. In some embodiments, the mobile device can capture an image of the sample while the sample is in the QMAX device and in the field of view (FOV) of a camera, in some embodiments, the camera is part of the mobile device.

[0399] In some embodiments, the adapter includes a plurality of optical components configured to enhance, amplify and / or optimize the generation of a signal from a sample. In some embodiments, the optical components include a plurality of portions configured to enhance, amplify and / or optimize the illumination provided to the sample. In certain embodiments, the illumination is provided by a light source that is part of the mobile device. In some embodiments, the optical components include a plurality of portions configured to enhance, amplify and / or optimize the signal from the sample. In some embodiments, the structure, function, and construction of the optical components can be found in PCT Application Nos. PCT / US2016 / 045437 (designating the United States) filed on August 10, 2016 and PCT / US0216 / 051775 filed on September 14, 2016, U.S. Provisional Application Nos. 62 / 456,590 filed on February 8, 2017, 62 / 459,554 filed on February 15, 2017, and 62 / 460,075 filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0400] 8. size

[0401] The devices, apparatus, systems and methods disclosed herein may include or use a QMAX device, which may include a plate and a spacer. In some embodiments, the dimensions of the various components of the QMAX device and its adapters are listed, described and / or summarized in PCT Application (designating the United States) No. PCT / US2016 / 045437 filed on August 10, 2016 and U.S. Provisional Application Nos. 62,431,639 filed on December 9, 2016 and 62 / 456,287 filed on February 8, 2017, which are hereby incorporated by reference in their entirety.

[0402] In some embodiments, the dimensions are listed in the following table:

[0403] plate:

[0404]

[0405]

[0406] Hinge:

[0407]

[0408]

[0409] Notch:

[0410]

[0411] Grooves:

[0412]

[0413]

[0414] Socket slot

[0415]

[0416] 9. Manual pressing

[0417] For the devices, apparatus, systems and methods disclosed herein, human hands can be used to manipulate or operate the plates and / or samples. In some embodiments, human hands can be used to press the plates into a closed configuration; in some embodiments, human hands can be used to press the samples into a thin layer. Methods of using hand pressure are described and / or summarized in PCT Applications (designating the United States) Nos. PCT / US2016 / 046437, filed on August 10, 2016, and PCT / US2016 / 051775, filed on September 14, 2016, and U.S. Provisional Applications Nos. 62 / 431,639, filed on December 9, 2016, 62 / 456,287, filed on February 8, 2017, 62 / 456,065, filed on February 7, 2017, 62 / 456,504, filed on February 8, 2017, and 62 / 460,062, filed on February 16, 2017, all of which are incorporated herein by reference.

[0418] In some embodiments, the plates of the QMAX device can be manipulated or operated by human hands. In certain embodiments, human hands can be used to apply an imprecise force to compress the plates from an open configuration to a closed configuration. In certain embodiments, human hands can be used to apply an imprecise force to achieve a high level of uniformity in sample thickness (e.g., less than 5%, 10%, 15%, or 20% variability).

[0419] In certain embodiments, the plates are movable relative to each other into different configurations, including an open configuration and a closed configuration; in the open configuration, the two plates are partially or completely separated, the spacing between the plates is not adjusted by spacers, and the sample is deposited on one or both of the plates; in the closed configuration: at least a portion of the sample is compressed by the two plates into a layer of very uniform thickness and is essentially stagnant relative to the plates, wherein the uniform thickness of the layer is defined by the sample contact area of ​​the two plates and adjusted by the plates and spacers; in some embodiments, the force pressing the two plates into the closed configuration is an imprecise pressing force provided by a human hand.

[0420] In some embodiments, the plates are conformally pressed. In some embodiments, the regions of at least one of the plates are conformally pressed in parallel or sequentially to press the plates together to form a closed configuration, wherein the conformal pressing produces a substantially uniform pressure on at least a portion of the sample of the plates, and the pressing causes at least a portion of the sample to expand laterally between the sample contacting surfaces of the plates, and wherein the closed configuration is a configuration in which the spacing between the plates in the uniform thickness region layer is adjusted by a spacer; in some embodiments, conformal pressing is a method of making the pressure applied to an area substantially constant regardless of changes in the shape of the outer surface of the plate. In some embodiments, parallel pressing applies pressure to the intended area simultaneously, and sequential pressing applies pressure to a portion of the intended area and gradually moves to other areas.

[0421] In some embodiments, the panels are pressed into a closed configuration by an imprecise force. In certain embodiments, the imprecise force is applied by a human hand. In some embodiments, the pressure is an imprecise force having a magnitude that is (a) unknown and unpredictable when the force is applied, or (b) unknown and cannot be predicted within an accuracy of 30% or better than the applied force. In some embodiments, the pressure is an imprecise force having a magnitude that cannot be determined within an accuracy of 30%, 40%, 50%, 70%, 100%, 200%, 300%, 500%, 1000%, 2000%, or any range between the two values ​​when the force is applied.

[0422] 10. Smartphone

[0423] The devices, apparatus, systems and methods disclosed herein may be used with mobile devices such as, but not limited to, smartphones. Smartphone detection techniques are disclosed herein or listed, described and summarized in PCT Applications (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, U.S. Provisional Application No. 62 / 456504, filed on February 8, 2017, all of which are incorporated herein in their entirety for all purposes.

[0424] In some embodiments, the smartphone includes a camera that can be used to capture an image or sample when the sample is in the camera's field of view (e.g., via an adapter). In some embodiments, the camera includes a set of lenses (e.g., an iPhone TM 6). In some embodiments, the camera includes at least two sets of lenses (e.g., iPhone TM 7) In some embodiments, the smartphone includes a camera, but the camera is not used for image capture.

[0425] In some embodiments, the smartphone includes a light source, such as, but not limited to, an LED (light emitting diode). In certain embodiments, the light source is used to provide illumination to the sample when the sample is in the field of view of the camera (e.g., through an adapter). In some embodiments, the light from the light source is enhanced, amplified, altered, and / or optimized by the optical components of the adapter.

[0426] In some embodiments, the smartphone includes a processor configured to process information from the sample. The smartphone includes software instructions that, when executed by the processor, can enhance, amplify and / or optimize the signal (e.g., image) from the sample. The processor may include one or more hardware components, such as a central processing unit (CPU), an application specific integrated circuit (ASIC), an application specific instruction set processor (ASIP), a graphics processing unit (GPU), a physical processing unit (PPU), a digital signal processor (DSP), a field programmable gate array (FPGA), a programmable logic device (PLD), a controller, a microcontroller unit, a reduced instruction set computer (RISC), a microprocessor, etc., or any combination thereof.

[0427] In some embodiments, the smart phone includes a communication unit configured and / or used to send data and / or images related to the sample to another device. By way of example only, the communication unit can use a cable network, a wired network, an optical fiber network, a telecommunications network, an intranet, the Internet, a local area network (LAN), a wide area network (WAN), a wireless local area network (WLAN), a metropolitan area network (MAN), a wide area network (WAN), a public switched telephone network (PSTN), a Bluetooth network, a ZigBee network, a near field communication (NFC) network, etc., or any combination thereof.

[0428] In some embodiments, the smartphone is an iPhone TM 、Android TM Phone or Windows TM Telephone.

[0429] 11. cloud

[0430] The devices, apparatus, systems and methods disclosed herein can be used with cloud storage and computing technologies. Relevant cloud technologies are disclosed or listed, described and summarized in PCT Applications (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.

[0431] In some embodiments, cloud storage and computing technology may involve a cloud database. By way of example only, a cloud platform may include a private cloud, a public cloud, a hybrid cloud, a community cloud, a distributed cloud, an inter-cloud, a multi-cloud, etc., or any combination thereof. In some embodiments, a mobile device (e.g., a smartphone) may be connected to the cloud via any type of network, including a local area network (LAN) or a wide area network (WAN).

[0432] In some embodiments, data associated with the sample (e.g., an image of the sample) is sent to the cloud without processing by the mobile device, and further analysis can be performed remotely. In some embodiments, data associated with the sample is processed by the mobile device, and the results are sent to the cloud. In some embodiments, both the raw data and the results are sent to the cloud.

Claims

1. A method for examining the effect of a test agent on one or more microbial individuals in a sample, the method comprising: (a) placing the test reagent and the detection reagent between two plates, wherein the test reagent is a fluorescent dye that labels individual microorganisms, and one or both plates have a spacer and one or both plates have a sample contact area, (b) Pressing the plate to form a uniform test layer (c) using an imager, obtaining a series of images of signals emanating from said individual microorganisms in said uniform test layer at at least two discrete time points; as well as (d) analyzing each image at the two discrete time points to determine changes in the individual microorganisms between the two discrete time points; wherein the two panels are movable relative to each other into different configurations, including an open configuration and a closed configuration, and at least one of the panels is a flexible panel; wherein the spacers have a spacer spacing ISD of 500 μm or less, the fourth power of the spacer spacing divided by the thickness h of the flexible board and the Young's modulus E, ISD 4 / (hE) equal to or less than 5x10 5 µm 3 / GPa, and the product of the thickness of the flexible plate and the Young's modulus is between 60 and 750 GPa-µm; 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; wherein in the closed configuration, which is configured after the sample is deposited in the open configuration: at least a portion of the sample is compressed by the two plates into the uniform test layer having a very uniform thickness, and the uniform thickness of the layer is defined by the sample contacting surfaces of the plates and adjusted by the plates and the spacers; wherein at least one of the spacers is in the sample contacting region; wherein said imaging is performed in a closed configuration, and wherein in said closed configuration said test layer thickness is configured such that there is substantially no overlap between any two microorganism individuals; and Wherein in the closed configuration, the thickness of the uniform test layer is adjusted by the two plates and the spacers in the sample area, and the height of the spacers is 250 microns or less.

2. A method for examining the effect of a test agent on one or more microbial individuals in a sample, the method comprising: (a) placing the test reagent and a sample containing one or more microorganisms between two plates to form a uniform test layer, (b) using an imager, obtaining at least two images of the uniform test layer at discrete points in time; (c) analyzing the image to determine changes in the individual microorganisms between at least two of the discrete time points; wherein the thickness of the uniform test layer is adjusted by two plates and spacers between the two plates and within the sample; wherein the thickness of the test layer is configured such that there is substantially no overlap between two microorganism individuals; wherein the two plates are movable relative to each other into different configurations, including an open configuration and a closed configuration; wherein each plate has a sample contact area; wherein in the open configuration, the two plates are partially or fully separated, the spacing between the two plates is not regulated by the spacer, and the sample is deposited on one or both plates; wherein in the closed configuration, the closed configuration is configured after the sample is deposited in the open configuration; at least a portion of the sample is compressed by the two plates into the uniform test layer having a very uniform thickness, and the uniform thickness of the layer is defined by the sample contacting surfaces of the plates and adjusted by the plates and the spacers; At least one of the two plates is a flexible plate; wherein the spacers have a spacer spacing ISD of 500 μm or less, the fourth power of the spacer spacing divided by the thickness h of the flexible board and the Young's modulus E, ISD 4 / (hE) equal to or less than 5x10 5 µm 3 / GPa, and the product of the thickness of the flexible plate and the Young's modulus is between 60 and 750 GPa-µm; At least one of the spacers is located in the sample contacting region.

3. The method according to claim 1 or 2, further comprising the step of staining the microorganisms with a cell viability dye indicating whether the stained cells are alive or dead.

4. The method according to claim 1 or 2, further comprising an additional reagent for staining the microorganism individuals.

5. The method of claim 4, wherein the detection reagent or the additional reagent or the test reagent is coated on one or both of the plates prior to depositing the sample.

6. The method of claim 1 or 2, wherein the one or more microorganisms are selected from the group consisting of bacteria, fungi, archaea, viruses, protists, microfauna, myxozoans, arthropods, crustaceans and microscopic nematodes.

7. The method of claim 1 or 2, wherein the one or more microorganisms are selected from a genus, and the genus is selected from the group consisting of: Bacillus, Bartonella, Bordetella, Borrelia, Brucella, Campylobacter, Chlamydia and Chlamydophila, Clostridium, Corynebacterium, Enterococcus, Escherichia, Francisella, Haemophilus, Helicobacter, Legionella, Leptospira, Listeria, Mycobacterium, Mycoplasma, Neisseria, Pseudomonas, Rickettsia, Salmonella, Shigella, Staphylococcus, Streptococcus, Treponema, Ureaplasma, Vibrio and Yersinia.

8. The method according to claim 1 or 2, wherein the one or more microorganisms are selected from the group consisting of: Bacillus anthracis, Bacillus cereus, Bartonella henselae, Bartonella quinquefasciatus, Bordetella pertussis, Borrelia burgdorferi, Borrelia garinii, Borrelia afzaleri, Borrelia relapsing fever, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacter jejuni, Chlamydia pneumoniae, Chlamydia trachomatis, Chlamydia psittaci, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Clostridium spp., Clostridium tetani, Corynebacterium diphtheriae, Enterococcus faecalis, Enterococcus faecium, Escherichia coli, Tularemia tularensis Francisella, Haemophilus influenzae, Helicobacter pylori, Legionella pneumophila, Leptospira interrogans, Leptospira sanguinis, Leptospira welchii, Leptospira noguchi, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium tuberculosis, Mycobacterium ulcerans, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis, Pseudomonas aeruginosa, Rickettsia spp., Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Treponema pallidum, Ureaplasma urealyticum, Vibrio cholerae, Yersinia pestis, Yersinia enterocolitica, and Yersinia pseudotuberculosis.

9. The method of claim 1 or 2, wherein the test agent is selected from the group consisting of: antibiotics, and antifungal agents.

10. The method of claim 1 or 2, wherein the test agent comprises an antibiotic, and wherein the antibiotic is selected from the group consisting of: actinomycin D, actinomycin, aculacin A, acyclovir, adenine 9-β-D-arabinofuranoside, alamethicin, L-alanine-L-1-aminoethylphosphonic acid, albendazole, 17-(allylamino)-17-demethoxygeldanamycin, astatin, amikacin, 7-aminoactinomycin D, 7-aminocephalosporanic acid, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, (+)-6-aminopenicillanic acid, amoxicillin, amphotericin B, ampicillin, anhydroerythromycin A, anisomycin, antimycin A, antimycin A2, antipain, aphidicolin, aphidicolin, apoptolidin A, Apramycin, Artesunate, Ascochlorin, Ascomycin, 5-Azacytidine, Azaserine, Azithromycin, Azlocillin, Bacitracin, Antimicrobial Peptides, Bafilomycin A1, Bafilomycin B1, Benstatin, βD-4 Human Recombinant, βD-1 (36 amino acids) Human Recombinant, βD-1 (47 amino acids) Human Recombinant, Bithionol VETRANAL™, Blasticidin S, Bleomycin, BM-Cyclin, Borrelidin, Brefeldin A, Penicillin A, Calcium Ionophore III Selector™, Calcium Ionophore A23187, (S)-(+)-Camptothecin, Capreomycin, Carbadox, Carbenicillin, Carboplatin, Cecropin A, Cecropin B, Cecropin P1, Cefaclor, Cephalexin VETRANAL™, Cefazolin, Cefixime, Cefmetazole, Cefoperazone, Cefsulodin, Ceftazidime, Ceftriaxone, Cephalexin, Cephalosporin, Cefotaxime, Cefradine, Cercosporin, Cerulenin, Cetylpyridine, Chloramphenicol, Chlorhexidine, Chloroquine , chlortetracycline, chromomycin A3, chrysomecin A, chrysomecin B, cinnamycin, cinoxacin, ciprofloxacin, clarithromycin, clindamycin, clofazimine, clotrimazole, cloxacillin, colistin, polyacetylneuraminic acid, compound 48 / 80, concanavalin A, cordycepin, kumarin A1, cryptotanshinone, cycloheximide, D-cycloserine, cyclosporin A, cyclosporin C, cytochalasin D, cytochalasin B, dacarbazine antitumor purine analogs, daunorubicin, 10-deacetylbaccatin III, dequamectin, defensin HNP-1, defensin HNP-2, demeclocycline hydrochloride, 1-deoxymannonojirimycin, 1-deoxynojirimycin, 6,9-Diamino-2-ethoxyacridine-DL-lactate, cis-diammineplatinum(II), 6-diazo-5-oxo-L-norleucine, 5,7-Dichloro-8-hydroxy-2-methylquinoline, Diclofenac PESTANAL®, Dicloxacillin, Diethylcarbamazine, Difloxacin, Dihydrostreptomycin, Diloxanide furoate, Dimethoprim, Dirithromycin, Doxorubicin, Doxycycline, Duramycin, Econazole, Elastin, Anthraquinone, Emetine, Enrofloxacin, Erythromycin, Ethambutol, Etoposide, Fengyuansu, Serine, Florfenicol, Flubendazole VETRANAL™, Fluconazole, Flumequine, Flumethasone, 5-fluorocytidine nucleoside analogs, Flurbiprofen Cyclooxygenase inhibitors, Metamycin A, Fumagillin, Fumagillin C, Furazolidone, Fusaric acid, G 418, ganciclovir, gatifloxacin, geldamycin, gentamicin, crystal violet, gliotoxin, gramicidin, griseofulvin, herbimycin A, cetylpyridine, magnolol, hydrocortisone 21, 8-hydroxyquinoline, 4-hydroxytamoxifen, (Z)-4-hydroxytamoxifen, hygromycin B, canthamicin, imipenem, indomethacin, ionomycin, Irgasan, istracycline Conazole, iturin A, ivermectin, josamycin, K-252a, K-252b, kanamycin, kasugamycin, kendomycin, ketoconazole, flavomycin, L-(+)-lactic acid, lactoferrin B, leptomycin A, leptomycin B, levamisole, levofloxacin, lincomycin, Listeria monocytogenes selective supplement I, LL-37, lomefloxacin, lysostaphin, bombesin I, mefenamic acid azole, meclocycline, menadione, 2-thiopyridine N-oxide, N-methyl-1-deoxynojirimycin, 2-methyl-4-isothiazoline-3-hydrochloride, metronidazole, mevastatin, (±)-miconazole, minocycline, mithramycin A, mitomycin C, monensin, morantel, hydroxycarboxylic acid oxadiazine, mupirocin, subtilisin, myxothiazole, propylthioamide, nafcillin, naftifine, nalidixic acid, methyl narasin , neocarcinostenin, neomycin, netilmicin, neterotine, niclosamide, nigericin, nikkomycin Z, nisin, nitrofurantoin, nogamycin, no active nicotinic acid, norfloxacin, nourseothricin, neomycin, NP-1, nystatin, ochratoxin A, ofloxacin, oligomycin, oligomycin A, neocyandiamide, octacycline, oxazolidinone, oxytetracycline, oxytetracycline, paclitaxel, paromomycin, patulin, PD 404,182, pediocin, pefloxacin, D-penicillamine, penicillin G, penicillin V, pentamidine, PGLa, 1,10-phenanthroline, phenazine, phleomycin, fosfomycin, pimaricin, pipemidic acid, piperacillin, pirarubicin, plate mycin, polymyxin B, poly(vinyl pyrrolidone) iodine complex, potassium clavulanate:cellulose 1:1, potassium sorbate, praziquantel, puromycin, pyrantel, pyrazinamide, bisquinacridone, pyrrolocin, quinine, 8-quinolinol, radicicol, ramoplanin, rapamycin, butterfly mycin, rafamycin A, ribavirin, ribostamycin, albendazole sulfoxide, rifabutin, rifampicin, rifamycin SV, rifapentine, rifaximin, ristocetin, rolicycline, roxithromycin, salinomycin , Sanmycin, Sinefungin, Perillomycin, Sorbic acid, Sordolin, Sparfloxacin, Spectinomycin, Spergualin, Spiramycin, Staurosporine, Streptolysin O, Streptomycin, Streptomycin, Streptozotocin, Succinylsulfathiazole, Sulconazole, Sulfadiazine, Sulfadiazine, Sulfadiazine, Sulfadimethoxine, Sulfadimethoxine, Sulfadiaz ... E, tamoxifen, tazobactam, teicoplanin, terbinafine, terconazole, tetracycline, tetramidazole, thiabendazole, thiamphenicol, sodium thimerosal, thiocandin, thiostrepton, thio-TEPA, thymol, tiamulin, hydroxythiophene penicillin, tioconazole, aminoglycoside antibiotics, tobramycin, tolnaftate, toyocamycin, triazine C, trichlorfon, trimethoprim, tuberculin, tunicamycin, tunicamycin C2 homologues, tylosin, valacyclovir, valinomycin, vinblastine, virginiamycin S1, virginiamycin M1, and analogs thereof, and salts thereof, and derivatives thereof. , 11. The method of claim 1 or 2, wherein the test agent comprises an antifungal agent, and wherein the antifungal agent is selected from the group consisting of: polyene antifungal agents imidazoles, triazoles, thiazoles, allylamines, echinocandins aurone, benzoic acid, ciclopirox, flucytosine and 5-fluorocytosine, griseofulvin, chloropropynyl iodide, tolnaftate, undecylenic acid, crystal violet, balsam peru, analogs thereof, salts thereof, and derivatives thereof.

12. The method of claim 1 or 2, wherein the imager comprises a camera.

13. The method of claim 1 or 2, wherein the imager comprises a camera.

14. The method of claim 1 or 2, wherein the uniform height of the spacers is equal to or less than an average size of the one or more microorganisms.

15. The method of claim 1 or 2, wherein the uniform height of the spacers is in the range of 75% to 125% of the average size of the one or more microorganisms.

16. The method of claim 1 or 2, wherein at least one of the plates is transparent.

17. The method of claim 1 or 2, wherein one or both of the plates contain a dye on the corresponding sample contact area that dissolves in the sample and stains the one or more microorganisms when in contact with the sample.

18. The method of claim 1 or 2, wherein the sample comprises an agent that promotes the survival and / or division of the one or more microorganisms.

19. The method of claim 1 or 2, wherein one or both of the plates contain reagents on the respective sample contact areas that promote survival and / or division of the microorganisms.

20. The method according to claim 1 or 2, further comprising: A housing is configured to hold the sample and to be mounted to the mobile communication device.

21. The method of claim 1 or 2, wherein the imager is configured to obtain a series of images of the microorganism at discrete time points.

22. The method of claim 1 or 2, further comprising a processor configured to receive and analyze the series of images, and identify and count the microorganisms in the first region in each of the images.

23. The method of claim 1 or 2, wherein the imager is configured to obtain a video comprising a series of images of the microorganism over a first period of time.

24. The method of claim 20, wherein the housing comprises optics for facilitating imaging and / or signal processing of the sample by a mobile communication device, and a base configured to hold the optics on the mobile communication device.

25. The method of claim 24, wherein the mobile communication device is configured to transmit the test results to medical personnel, a medical institution, or an insurance company.

26. The method of claim 25, wherein the mobile communication device is further configured to communicate information about the subject with the medical personnel, medical institution, or insurance company.

27. The method of claim 24, wherein the mobile communication device is configured to receive a prescription, diagnosis, or advice from a medical professional.

28. The method of claim 24, wherein the mobile communication device communicates with a remote location via WiFi or a cellular network.

29. The method of claim 24, wherein the mobile communication device is a mobile phone.

30. The method of claim 1 or 2, wherein the one or more microorganisms have an average size of less than 1 mm.

31. A method according to claim 1 or 2, wherein one or both of the plates include a control zone and an experimental zone on the corresponding sample contact area, wherein the experimental zone contains one or more test reagents that dissolve and diffuse in the sample when contacted with the sample, and wherein the control zone does not contain such test reagents.

32. The method of claim 1 or 2, further comprising contacting the sample with an additional reagent, wherein the additional reagent is selected from the group consisting of: a staining dye that labels the microorganism, a life support reagent that promotes the survival of the microorganism, and a proliferation reagent that maintains or promotes the proliferation of the microorganism.

33. The method of claim 32, wherein the additional reagent comprises a staining dye comprising a fluorescent molecule selected from the group consisting of: IRDye800CW, Alexa 790, Dylight 800, fluorescein, fluorescein isothiocyanate, succinimidyl ester of carboxyfluorescein, succinimidyl ester of fluorescein, 5-isomer of fluorescein dichlorotriazine, caged carboxyfluorescein-alanine-carboxamide, Oregon Green 488, Oregon Green 514; fluorescein yellow, acridine orange, rhodamine, Texas Red, propidium iodide, JC-1 (5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide), tetrabromorhodamine 123, tetramethylrhodamine methyl ester, tetramethylrhodamine ethyl ester, tetramethylrhodamine, 4- Dimethylaminotetramethylrhodamine, green fluorescent protein, blue-shifted green fluorescent protein, blue-green-shifted green fluorescent protein, red-shifted green fluorescent protein, yellow-shifted green fluorescent protein, 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid; acridine and derivatives; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid (EDANS); 4-amino-N-[(3-vinylsulfonyl)phenyl]naphthalenamide-3,5-disulfonate; N-(4-anilino-1-naphthyl)maleimide; o-aminobenzamide; 4,4-difluoro-5-(2 -thienyl)-4-boron-3a,4a-diaza-5-indacene-3-propionic acid BODIPY; Cascade Blue; Brilliant Yellow; Coumarin and derivatives: Coumarin, 7-amino-4-methylcoumarin), 7-amino-4-methylcoumarin; Cyanine dyes; Labeling Red; 4',6-diamino-2-phenylindole; 5',5''-dibromopyrogallolsulfonaphthalene (bromopyrogallol red); 7-diethylamino-3-(4'-isothiocyanatophenyl)-4-methylcoumarin; Diethylenetriaminepentaacetate; 4,4'-diisothiocyanatostilbene-2,2' -disulfonic acid; 4,4'-diisothiocyanato-2,2'-yldisulfonic acid disodium salt; 5-(dimethylamino)naphthalene-1-sulfonyl chloride (DNS, dansyl chloride); 4-dimethylaminophenylazophenyl-4'-isothiocyanate; eosin and derivatives: eosin, eosin isothiocyanate, erythrosine and derivatives: erythrosine B, erythrosine, isothiocyanate; ethidium; fluorescein and its derivatives: 5-carboxyfluorescein, 5-(4,6-dichlorotriazin-2-yl)amino-fluorescein (DTAF), 2',7'-dimethoxy-4',5'-dichloro-6-carboxyfluorescein, Fluorescein isothiocyanate, QFITC, XRITC; fluorescamine; IR144; IR1446; malachite green isothiocyanate; 4-methylumbelliferyl o-cresolphthalein; nitrotyrosine; para-fuchsin; phenol red; B-phycoerythrin; o-phthalaldehyde; pyrene and its derivatives: pyrene, pyrenebutyric acid, succinimidyl 1-pyrene; butyrate quantum dots;Reactive Red 4 Rhodamine and derivatives: 6-Carboxy-X-Rhodamine, 6-Carboxyrhodamine, Lissamine Rhodamine B Sulfonyl Chloride Rhodamine, Rhodamine B, Rhodamine 123, Rhodamine X Isothiocyanate, Sulforhodamine B, Sulforhodamine 101, Sulfonyl Chloride Derivatives of 5-Sulforhodamine; N,N,N',N'-Tetramethyl-6-Carboxyrhodamine; Tetramethylrhodamine; Tetramethylrhodamine isothiocyanate; Riboflavin; 5-(2'-aminoethyl)aminonaphthalene-1-sulfonic acid), 4-(4'-dimethylaminophenylazo)benzoic acid rhodamine; CAL Fluorescent Orange 560; Terbium Chelate Derivatives; Cy3; Cy5; Cy5.5; Cy7; IRD700; IRD 800; La Jolla blue; phthalocyanines; and naphthalocyanines, coumarins and related dyes, xanthene dyes, resorufins, bimanes acridines, isoindole, dansyl dyes, aminophthalhydrazides and isoluminol derivatives, aminophthalimides, aminonaphthalimides, aminobenzofurans, aminoquinolines, dicyanohydroquinones, fluorescent europium and terbium complexes; suitable fluorescent and chromogenic proteins include, but are not limited to, green fluorescent proteins, including, but not limited to, GFP derived from Aequoria victoria or a derivative thereof; GFP from another species, including Renilla müller or Ptilosarcus guernyi; humanized recombinant GFP; any of a variety of fluorescent and colored proteins from coral species; and any combination thereof. ; 34. The method of claim 1 or 2, wherein it further comprises a life support agent that promotes the survival of microorganisms.

35. The method of claim 11, wherein The polyene antifungal agent is selected from the group consisting of amphotericin B, candidiasis, filipinocin, harvesin, natamycin, nystatin, cytomycin or a combination thereof, The imidazole is selected from the group consisting of bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole or a combination thereof; The triazole is selected from the group consisting of albaconazole, efenaconazole, epoxiconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, ravuconazole, terconazole, voriconazole or a combination thereof; The thiazole refers to abafungin; The allylamine is selected from the group consisting of amorolfine, butenafine, naftifine and terbinafine or a combination thereof; The echinocandin is selected from the group consisting of anidulafungin, caspofungin, micafungin or a combination thereof.

36. The method of claim 33, wherein the staining dye comprises a cell viability dye, The life support agent comprises nutrients, an oxygen source, a pH buffer, or a combination thereof; The proliferation agent includes hormones, iron carriers or a combination thereof.

37. The method of claim 32, wherein the additional reagent comprises a dye selected from the group consisting of fluorescent proteins, chromogenic proteins, and combinations thereof.

38. The method of claim 37, wherein the fluorescent protein is green fluorescent protein (GFP).

39. The method of claim 38, wherein the GFP is GFP derived from Aequorea victoria.

40. The method of claim 38, wherein the GFP is humanized or enhanced GFP.

41. The method of claim 38, wherein the GFP is from Renilla or Renilla glauca.

42. The method of claim 37, wherein the fluorescent protein or chromogenic protein is from a coral polyp species.

43. The method of claim 1 or 2, further comprising contacting the sample with an additional reagent, wherein the additional reagent comprises a cell viability dye.

44. The method according to claim 1 or 2, wherein the two plates are connected by a hinge that allows the two plates to rotate.

45. The method of claim 1 or 2, wherein the spacer has a height in the range of 1 μm to 100 μm.

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