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WO2018152421A1 - Dosage pour condensats de vapeur - Google Patents

Dosage pour condensats de vapeur Download PDF

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Publication number
WO2018152421A1
WO2018152421A1 PCT/US2018/018520 US2018018520W WO2018152421A1 WO 2018152421 A1 WO2018152421 A1 WO 2018152421A1 US 2018018520 W US2018018520 W US 2018018520W WO 2018152421 A1 WO2018152421 A1 WO 2018152421A1
Authority
WO
WIPO (PCT)
Prior art keywords
prior
sample
plates
plate
spacers
Prior art date
Application number
PCT/US2018/018520
Other languages
English (en)
Inventor
Stephen Y. Chou
Ji QI
Wei Ding
Yufan ZHANG
Original Assignee
Essenlix Corporation
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from PCT/US2018/017307 external-priority patent/WO2018148342A1/fr
Priority claimed from PCT/US2018/017502 external-priority patent/WO2018148470A1/fr
Priority claimed from PCT/US2018/017489 external-priority patent/WO2018148458A1/fr
Priority claimed from PCT/US2018/017494 external-priority patent/WO2018148463A1/fr
Priority claimed from PCT/US2018/017504 external-priority patent/WO2018148471A2/fr
Priority claimed from PCT/US2018/017492 external-priority patent/WO2018148461A1/fr
Priority claimed from PCT/US2018/017501 external-priority patent/WO2018148469A1/fr
Priority claimed from PCT/US2018/017713 external-priority patent/WO2018148607A1/fr
Priority claimed from PCT/US2018/017716 external-priority patent/WO2018148609A2/fr
Priority claimed from PCT/US2018/017712 external-priority patent/WO2018148606A1/fr
Priority claimed from PCT/US2018/018007 external-priority patent/WO2018148729A1/fr
Priority claimed from PCT/US2018/018108 external-priority patent/WO2018148764A1/fr
Priority claimed from PCT/US2018/017499 external-priority patent/WO2018152005A1/fr
Priority claimed from PCT/US2018/018405 external-priority patent/WO2018152351A1/fr
Priority to US16/485,126 priority Critical patent/US11523752B2/en
Application filed by Essenlix Corporation filed Critical Essenlix Corporation
Publication of WO2018152421A1 publication Critical patent/WO2018152421A1/fr
Priority to US17/980,400 priority patent/US20230077906A1/en

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B5/00—Measuring for diagnostic purposes; Identification of persons
    • A61B5/08—Measuring devices for evaluating the respiratory organs
    • A61B5/097—Devices for facilitating collection of breath or for directing breath into or through measuring devices

Definitions

  • PCT/US18/18007 filed on 2/13/2018 (18F1 1 B), PCT Application No. PCT/US18/17716, filed on 2/9/2018 (18F08), PCT Application No. PCT/US18/17713, filed on 2/9/2018 (18F07), PCT Application No. PCT/US18/17712, filed on 2/9/2018 (18F16), PCT Application No.
  • PCT/US18/17504 filed on 2/8/2018 (18F02), PCT Application No. PCT/US18/17501 , filed on 2/8/2018 (18F12), PCT Application No. PCT/US18/17499, filed on 2/8/2018 (18F17), PCT Application No. PCT/US18/17489, filed on 2/8/2018 (18F18), PCT Application No.
  • PCT/US18/17492 filed on 2/8/2018 (18F10)
  • PCT Application No. PCT/US18/17494 filed on 2/8/2018 (18F02
  • PCT Application No. PCT/US18/17502 filed on 2/8/2018 (18F09
  • PCT Application No. PCT/US18/17307 filed on 2/7/2018 (18F15A)
  • the present invention is related to the field of bio/chemical sampling, sensing, assays and applications.
  • the present invention relates to provide, among other things, the methods, devices, and systems that can address these needs.
  • the present invention is related to provide, among other things, the device, apparatus, systems, and methods for:
  • a vapor condensate sample e.g. the exhaled breath condensate (EBC) from a subject
  • the devices can get the sample thickness information either using spacers or without a spacers.
  • EBC exhaled breath condensate
  • Fig. 1 An illustration of certain aspects of an exemplary device and methods of collecting exhaled breath condensate (EBC) using a SiEBCA (Single-drop EBC
  • Collector/Analyzer that comprises a pair of movable plates without using spacers.
  • FIG. 2 An illustration of certain aspects of an exemplary device and methods of collecting exhaled breath condensate (EBC) using a SiEBCA (Single-drop EBC
  • FIG. 3 An illustration of different formations of EBC at closed configuration of SiEBCA depends on spacer height.
  • closed configuration- 1 If spacer height is smaller than average height of EBC at open configuration; at closed configuration, EBC become a continuous thin film contacting both collection and cover plates and may have air isolated pockets.
  • closed condiguraiton-2 If spacer height is larger than average height of EBC at open configuration; at closed configuration EBC become isolated puddle(s) that contact both collection and cover plates, and that are larger but fewer than that at the open configuration.
  • Fig. 4 An illustration of an embodiment of the devices and the methods of a SiEBCA (Single-drop EBC Collector/Analyzer).
  • Fig. 5a An illustration of a SiEBCA with both "open spacer” and “enclosed spacer", where the open spacer is a post (pillar) while the enclosed spacer is a ring spacer (4) and a four-chamber grid spacer (5).
  • Fig. 5b shows reducing binding or mixing time by reducing the sample thickness using two pates, spacers, and compression (shown in cross-section).
  • Panel (1) illustrates reducing the time for binding entities in a sample to a binding site on a solid surface (X-(Volume to Surface)).
  • Panel (2) illustrates reducing the time for binding entities (e.g. reagent) stored on a surface of plate to a binding site on a surface of another surface (X- (Surface to Surface)).
  • Panel (3) illustrates reducing the time for adding reagents stored on a surface of a plate into a sample that is sandwiched between the plate and other plate (X- (Surface to Volume)).
  • Fig. 6 schematically illustrates the optical measurement and imaging taken for the measurement of the EBC sample thickness and lateral area, respectively.
  • Fig. 7 demonstrates the principle of plate spacing measurement based on F-P cavity resonance.
  • Fig. 8 shows microscopic images of the EBC sample collected using the exemplary SiEBCA device without spacers.
  • EBC exhaled breath condensate
  • VC vapor condensate
  • SiEBCA and "SiEBC” are interchangeable.
  • polynucleotide refers to a binding member, e.g.
  • nucleic acid molecule nucleic acid molecule, polypeptide molecule, or any other molecule or compound, that can specifically bind to its binding partner, e.g., a second nucleic acid molecule containing nucleotide sequences complementary to a first nucleic acid molecule, an antibody that specifically recognizes an antigen, an antigen specifically recognized by an antibody, a nucleic acid aptamer that can specifically bind to a target molecule, etc.
  • a secondary capture agent which can also be referred to as a “detection agent” refers a group of biomolecules or chemical compounds that have highly specific affinity to the antigen.
  • the secondary capture agent can be strongly linked to an optical detectable label, e.g., enzyme, fluorescence label, or can itself be detected by another detection agent that is linked to an optical detectable label through bioconjugation (Hermanson, "Bioconjugate Techniques” Academic Press, 2nd Ed., 2008).
  • capture agent-reactive group refers to a moiety of chemical function in a molecule that is reactive with capture agents, i.e., can react with a moiety (e.g., a hydroxyl, sulfhydryl, carboxyl or amine group) in a capture agent to produce a stable strong, e.g., covalent bond.
  • a moiety e.g., a hydroxyl, sulfhydryl, carboxyl or amine group
  • specific binding and “selective binding” refer to the ability of a capture agent to preferentially bind to a particular target analyte that is present in a heterogeneous mixture of different target analytes.
  • a specific or selective binding interaction will discriminate between desirable (e.g., active) and undesirable (e.g., inactive) target analytes in a sample, typically more than about 10 to 100-fold or more (e.g., more than about 1000- or 10,000-fold).
  • analyte refers to a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecule), cells, tissues, viruses, and nanoparticles with different shapes.
  • test refers to testing a sample to detect the presence and/or abundance of an analyte.
  • determining As used herein, the terms “determining,” “measuring,” and “assessing,” and “assaying” are used interchangeably and include both quantitative and qualitative determinations.
  • the term "light-emitting label” refers to a label that can emit light when under an external excitation. This can be luminescence. Fluorescent labels (which include dye molecules or quantum dots), and luminescent labels (e.g., electro- or chemi-luminescent labels) are types of light-emitting label.
  • the external excitation is light (photons) for fluorescence, electrical current for electroluminescence and chemical reaction for chemi-luminescence. An external excitation can be a combination of the above.
  • labeled analyte refers to an analyte that is detectably labeled with a light emitting label such that the analyte can be detected by assessing the presence of the label.
  • a labeled analyte may be labeled directly (i.e., the analyte itself may be directly conjugated to a label, e.g., via a strong bond, e.g., a covalent or non-covalent bond), or a labeled analyte may be labeled indirectly (i.e., the analyte is bound by a secondary capture agent that is directly labeled).
  • hybridizing and “binding”, with respect to nucleic acids, are used
  • capture agent/analyte complex is a complex that results from the specific binding of a capture agent with an analyte.
  • a capture agent and an analyte for the capture agent will usually specifically bind to each other under "specific binding conditions” or “conditions suitable for specific binding”, where such conditions are those conditions (in terms of salt concentration, pH, detergent, protein concentration, temperature, etc.) which allow for binding to occur between capture agents and analytes to bind in solution.
  • specific binding conditions or “conditions suitable for specific binding”
  • Such conditions particularly with respect to antibodies and their antigens and nucleic acid hybridization are well known in the art (see, e.g., Harlow and Lane (Antibodies: A Laboratory Manual Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. (1989) and Ausubel, et al, Short Protocols in Molecular Biology, 5th ed., Wiley & Sons, 2002).
  • a subject may be any human or non-human animal.
  • a subject may be a person performing the instant method, a patient, a customer in a testing center, etc.
  • a "diagnostic sample” refers to any biological sample that is a bodily byproduct, such as bodily fluids, that has been derived from a subject.
  • the diagnostic sample may be obtained directly from the subject in the form of liquid, or may be derived from the subject by first placing the bodily byproduct in a solution, such as a buffer.
  • exemplary diagnostic samples include, but are not limited to, saliva, serum, blood, sputum, urine, sweat, lacrima, semen, feces, breath, biopsies, mucus, etc.
  • an "environmental sample” refers to any sample that is obtained from the environment.
  • An environmental sample may include liquid samples from a river, lake, pond, ocean, glaciers, icebergs, rain, snow, sewage, reservoirs, tap water, drinking water, etc.; solid samples from soil, compost, sand, rocks, concrete, wood, brick, sewage, etc.; and gaseous samples from the air, underwater heat vents, industrial exhaust, vehicular exhaust, etc.
  • samples that are not in liquid form are converted to liquid form before analyzing the sample with the present method.
  • a "foodstuff sample” refers to any sample that is suitable for animal consumption, e.g., human consumption.
  • a foodstuff sample may include raw ingredients, cooked food, plant and animal sources of food, preprocessed food as well as partially or fully processed food, etc.
  • samples that are not in liquid form are converted to liquid form before analyzing the sample with the present method.
  • diagnosis refers to the use of a method or an analyte for identifying, predicting the outcome of and/or predicting treatment response of a disease or condition of interest.
  • a diagnosis may include predicting the likelihood of or a predisposition to having a disease or condition, estimating the severity of a disease or condition, determining the risk of progression in a disease or condition, assessing the clinical response to a treatment, and/or predicting the response to treatment.
  • a “biomarker,” as used herein, is any molecule or compound that is found in a sample of interest and that is known to be diagnostic of or associated with the presence of or a
  • Biomarkers include, but are not limited to, polypeptides or a complex thereof (e.g., antigen, antibody), nucleic acids (e.g., DNA, miRNA, mRNA), drug metabolites, lipids, carbohydrates, hormones, vitamins, etc., that are known to be associated with a disease or condition of interest.
  • polypeptides or a complex thereof e.g., antigen, antibody
  • nucleic acids e.g., DNA, miRNA, mRNA
  • drug metabolites lipids, carbohydrates, hormones, vitamins, etc.
  • a “condition” as used herein with respect to diagnosing a health condition refers to a physiological state of mind or body that is distinguishable from other physiological states.
  • a health condition may not be diagnosed as a disease in some cases.
  • Exemplary health conditions of interest include, but are not limited to, nutritional health; aging; exposure to environmental toxins, pesticides, herbicides, synthetic hormone analogs; pregnancy;
  • entity refers to, but not limited to proteins, peptides, DNA, RNA, nucleic acid, molecules (small or large), cells, tissues, viruses, nanoparticles with different shapes, that would bind to a "binding site".
  • entity includes the capture agent, detection agent, and blocking agent.
  • the “entity” includes the “analyte”, and the two terms are used interchangeably.
  • binding site refers to a location on a solid surface that can immobilize an entity in a sample.
  • entity partners refers to, but not limited to proteins, peptides, DNA, RNA, nucleic acid, molecules (small or large), cells, tissues, viruses, nanoparticles with different shapes, that are on a "binding site” and would bind to the entity.
  • entity include, but not limited to, capture agents, detection agents, secondary detection agents, or "capture agent/analyte complex”.
  • smart phone or “mobile phone”, which are used interchangeably, refers to the type of phones that has a camera and communication hardware and software that can take an image using the camera, manipulate the image taken by the camera, and communicate data to a remote place.
  • the Smart Phone has a flash light.
  • average linear dimension of an area is defined as a length that equals to the area times 4 then divided by the perimeter of the area.
  • the area is a rectangle, that has width w, and length L, then the average of the linear dimension of the rectangle is
  • the average line dimension is, respectively, W for a square of a width W, and d for a circle with a diameter d.
  • the area includes, but not limited to, the area of a binding site or a storage site.
  • periodic structure array refers to the distance from the center of a structure to the center of the nearest neighboring identical structure.
  • the term "storage site” refers to a site of an area on a plate, wherein the site contains reagents to be added into a sample, and the reagents are capable of being dissolving into the sample that is in contract with the reagents and diffusing in the sample.
  • relevant means that it is relevant to detection of analytes, quantification and/or control of analyte or entity in a sample or on a plate, or quantification or control of reagent to be added to a sample or a plate.
  • hydrophilic means that the contact angle of a sample on the surface is less than 90 degree.
  • hydrophobic means that the contact angle of a sample on the surface is equal to or larger than 90 degree.
  • variable of a quantity refers to the difference between the actual value and the desired value or the average of the quantity.
  • relative variation refers to the ratio of the variation to the desired value or the average of the quantity. For example, if the desired value of a quantity is Q and the actual value is (Q+ ⁇ ), then the ⁇ /(Q+A) is the relative variation.
  • relative sample thickness variation refers to the ratio of the sample thickness variation to the average sample thickness.
  • optical transparent refers to a material that allows a transmission of an optical signal
  • optical signal refers to, unless specified otherwise, the optical signal that is used to probe a property of the sample, the plate, the spacers, the scale-marks, any structures used, or any combinations of thereof.
  • sample-volume refers to, at a closed configuration of a CROF process, the volume between the plates that is occupied not by the sample but by other objects that are not the sample.
  • the objects include, but not limited to, spacers, air bubbles, dusts, or any combinations of thereof. Often none-sample-volume(s) is mixed inside the sample.
  • saturation incubation time refers to the time needed for the binding between two types of molecules (e.g. capture agents and analytes) to reach an equilibrium.
  • the “saturation incubation time” refers the time needed for the binding between the target analyte (entity) in the sample and the binding site on plate surface reaches an equilibrium, namely, the time after which the average number of the target molecules (the entity) captured and immobilized by the binding site is statistically nearly constant.
  • first plate and “collection plate are interchangeable.
  • second plate and “cover plate” are interchangeable.
  • a “processor,” “communication device,” “mobile device,” refer to computer systems that contain basic electronic elements (including one or more of a memory, input-output interface, central processing unit, instructions, network interface, power source, etc.) to perform
  • the computer system may be a general purpose computer that contains instructions to perform a specific task, or may be a special-purpose computer.
  • a “site” or “location” as used in describing signal or data communication refers to the local area in which a device or subject resides.
  • a site may refer to a room within a building structure, such as a hospital, or a smaller geographically defined area within a larger
  • a remote site or remote location with reference to a first site that is remote from a second site, is a first site that is physically separated from the second site by distance and/or by physical obstruction.
  • the remote site may be a first site that is in a separate room from the second site in a building structure, a first site that is in a different building structure from the second site, a first site that is in a different city from the second site, etc.
  • sample collection site refers to a location at which a sample may be obtained from a subject.
  • a sample collection site may be, for example, a retailer location (e.g., a chain store, pharmacy, supermarket, or department store), a provider office, a physician's office, a hospital, the subject's home, a military site, an employer site, or other site or combination of sites.
  • sample collection site may also refer to a proprietor or representative of a business, service, or institution located at, or affiliated with, the site.
  • raw data includes signals and direct read-outs from sensors, cameras, and other components and instruments which detect or measure properties or characteristics of a sample.
  • Process management refers to any number of methods and systems for planning and/or monitoring the performance of a process, such as a sample analysis process.
  • One aspect of the present invention is that we developed and experimentally
  • VC vapor condensate
  • Exhaled breath condensate (EBC) analysis is a noninvasive method of detecting biomarkers, mainly coming from the lower respiratory tract. EBC is collected during quiet breathing, as a product of cooling and condensation of the exhaled aerosol.
  • An exemplary device and method of collecting exhaled breath condensate (EBC) using a SiEBC (single-drop exhaled breath condensate collector /analyzer) or SiVC (single-drop vapor condensate collector/analyzer) that does not have spacers, as illustrated in Fig. 1 , comprises the basic steps:
  • FIG. 1-1 A subject (e.g. human) breathe onto a plate, termed “collection plate”, and the breath condenses into EBC, which are in droplets with different sizes, depending on the breathing time. For a short breathing time most droplets are separated from each other.
  • the surface of the collection plate that collects the EBC is termed the sample surface; (2) placing a cover plate over the collection plate and pressing them together (Fig. 1- 2).
  • a cover plate with spacers (which are used for regulating the spacing between the cover plate and the substrate plate) is placed on top of the sample surface; and
  • the initial droplets are pressed into a thin layer EBC of a thickness that is regulated by the plates and spacers (not shown).
  • SiEBCA single drop
  • the substrate plate and “the cover plate” are respectively interchangeable with “the first plate” and “the second plate”.
  • the plates are cooled to reduce the evaporation of collected EBC.
  • a device for collecting and analyzing vapor condensate (VC) sample comprising: a collection plate and a cover plate, wherein:
  • the plates are movable relative to each other into different configurations
  • each of the plates has, on its inner respective surface, a sample contact area for contacting a vapor condensate (VC) sample that contains an analyte;
  • VC vapor condensate
  • one of the configurations is an open configuration, in which: the two plates are either completely or partially separated apart, and the VC sample is deposited on one or both of the plates;
  • another of the configurations is a closed configuration which is configured after the VC sample deposition in the open configuration; and in the closed configuration: at least a part of the VC sample is between the two plates and in contact with the two plates, and has a thickness that (a) is regulated by the two sample contact surfaces of the plates without using spacers, and (b) is equal to or less than 30 ⁇ with a small variation.
  • AAO-2 Another device is provided herein for collecting and analyzing vapor condensate
  • (VC) sample comprising:
  • a collection plate and a cover plate wherein: i. the plates are movable relative to each other into different configurations;
  • one or both plates are flexible
  • each of the plates has, on its respective surface, a sample contact area for
  • VC vapor condensate
  • one of the configurations is an open configuration, in which: the two plates are either completely or partially separated apart, and the VC sample is deposited on one or both of the plates;
  • another of the configurations is a closed configuration which is configured after the VC sample deposition in the open configuration; and in the closed configuration: at least a part of the VC sample is between the two plates and in contact with the two plates, and has a thickness that is regulated by the plate spacing without using spacers.
  • the thickness of the sample in a closed configuration of the plates is measured.
  • the sample thickness in a plate closed configuraiton include Fabry-Perot interferometer, or different optical focusing methods, or others.
  • an exemplary SiEBCA device comprises a collection plate of 25 mm X 25 mm X 1 mm PMMA planar plate with untreated surfaces, and a cover plate of 25 mm X 25 mm X 0.175 mm PMMA planar plate with bare untreated surfaces.
  • the EBC sample was collected by having a subject breathe on a collection plate for 2 sec and a cover plate was immediately brought to cover the collection plate and pressed against it as described above. Later, the SiEBCA together with the sample collected therein were subject to optical measurement and microscopy imaging.
  • Fig. 6 schematically illustrates the optical measurement and imaging taken for the measurement of the EBC sample thickness and lateral area, respectively.
  • panel (A) Fabry-Perot interferometer was used to measure the F-P cavity resonance in the reflectance spectra at 25 points on the 4X4 grid artificially generated in the center of the SiEBCA device, from which the plate spacing (and the sample thickness) is thus deduced.
  • Each of the 25 measuring points is about 2 um by 2 um in area, and all 25 points cover an area of 20 mm by 20 mm.
  • An average plate spacing over the 25 points was taken as the estimate of the sample thickness (f ⁇ ) .
  • the percentage of EBC liquid lateral area (a,) for each measuring point is calculated as (Si-Sb)/Si X 100%; second, an average value
  • the volume of the EBC sample ( VEBC) is thus determined as S EBC * T ⁇ C .
  • Fig. 7 demonstrates the principle of plate spacing measurement based on F-P cavity resonance.
  • Panel (a) shows the schematic of F-P cavity from the SiEBCA device; panel
  • h is the plate spacing
  • c is light speed
  • Av is the period in frequency domain
  • n is the reflective index of the EBC liquid.
  • the average EBC sample thickness is equal to—— .
  • Fig. 8 shows microscopic images of the EBC sample collected using the exemplary SiEBCA device without spacers.
  • Panels (a) - (b) respectively show the images of the EBC samples at the closed configuration after hand pressing the two plates with low, medium, and high pressing strength.
  • Low strength was less than 10 kg
  • high strength was higher than 15 kg
  • medium strength was in between the low and high strength.
  • a device for collecting and analyzing vapor condensate (VC) sample, comprising:
  • the plates are movable relative to each other into different configurations
  • one or both plates are flexible
  • each of the plates has, on its respective inner surface, a sample contact area for contacting a vapor condensate (VC) sample that contains an analyte;
  • VC vapor condensate
  • the spacers are fixed to the respective inner surface of one or both of the plates, and have a predetermined substantially uniform height and a predetermined constant inter-spacer distance and wherein at least one of the spacers is inside the sample contact area;
  • one of the configurations is an open configuration, in which: the two plates are either completely or partially separated apart, the spacing between the plates is not regulated by the spacers, and the VC sample is deposited on one or both of the plates;
  • another of the configurations is a closed configuration which is configured after the VC sample deposition in the open configuration; and in the closed configuration: at least a part of the VC sample is between the two plates and in contact with the two plates, and has a highly uniform thickness that is regulated by the spacers and the two sample contact surfaces of the plates and is equal to or less than 30 ⁇ .
  • the device further comprises, on one or both plates, one or a plurality of dry binding sites and/or one or a plurality of reagent sites.
  • the sample is exhale breath condensate.
  • the sample is a vapor from a biological sample, an environmental sample, a chemical sample, or clinical sample.
  • the analyte comprises a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecules), cells, tissues, viruses, and nanoparticles with different shapes.
  • the analyte comprises volatile organic compounds (VOCs).
  • VOCs volatile organic compounds
  • the analyte comprises nitrogen, oxygen, C02, H20, and inert gases.
  • the analyte is stained.
  • the device may comprise a dry reagent coated on one or both of the plates.
  • the dry reagent may bind to an analyte in the blood an immobilize the analyte on a surface on one or both of the plates.
  • the reagent may be an antibody or other specific binding agent, for example. This dry reagent may have a pre-determined area.
  • the device may comprise a releasable dry reagent on one or more of the plates, e.g., a labeled reagent such as a cell stain or a labeled detection agent such as an antibody or the like.
  • there may be a release time control material on the plate that contains the releasable dry reagent wherein the release time control material delays the time that the releasable dry regent is released into the blood sample.
  • the release time control material delays the time that the dry regent is released into the blood sample by at least 3 seconds, e.g., at least 5 seconds or at least 10 seconds.
  • the drive may contain multiple dry binding sites and/or multiple reagent sites, thereby allowing multiplex assays to be performed.
  • the areas occupied by the drying binding sites may oppose the areas occupied by the reagent sites when the plates are in the closed position.
  • the regent comprises labeling or staining reagent(s).
  • the spacers regulating the layer of uniform thickness i.e., the spacers that are spacing the plates away from each other in the layer
  • the Young's modulus of the spacers times the filling factor of the spacers is equal or larger than 10 MPa, e.g., at least 15 MPa or at least 20 MPa, where the filling factor is the ratio of the spacer area that is in contact with the layer of uniform thickness to the total plate area that is in contact with the layer of uniform thickness.
  • the thickness of the flexible plate times the Young's modulus of the flexible plate is in the range 60 to 750 GPa-um, e.g., 100 to 300 GPa-um, 300 to 550 GPa-um, or 550 to 750 GPa-um.
  • the fourth power of the inter-spacer-distance (ISD) divided by the thickness of the flexible plate (h) and the Young's modulus (E) of the flexible plate, ISD4/(hE), is equal to or less than 10 ⁇ 6 um3/GPa, e.g., less than 10 ⁇ 5 um3/GPa, less then 10M um3/GPa or less than 10 ⁇ 3 um3/GPa.
  • one or both plates comprises a location marker either on a surface of or inside the plate, that provide information of a location of the plate, e.g., a location that is going to be analyzed or a location onto which the blood should be deposited.
  • one or both plates may comprise a scale marker, either on a surface of or inside the plate, that provides information of a lateral dimension of a structure of the blood sample and/or the plate.
  • one or both plates comprises an imaging marker, either on surface of or inside the plate that assists an imaging of the sample.
  • the imaging marker could help focus the imaging device or direct the imaging device to a location on the device.
  • the spacers can function as a location marker, a scale marker, an imaging marker, or any combination of thereof.
  • it further comprises an enclosure- spacer that encloses a partial or entire VC samples deposited on the collection plate.
  • the highly uniform thickness has a value equal to or less than 0.5 um. In some embodiments, the highly uniform thickness has a value in the range of 0.5 um to 1 um, 1 um to 2 um, 2 um to 10 um, 10 um to 20 um or 20 um to 30 um.
  • the thickness of the at least a part of VC sample at the closed configuration is larger than the thickness of VC sample deposited on the collection plate at an open configuration.
  • the thickness of the at least a part of VC sample at the closed configuration is less than the thickness of VC sample deposited on the collection plate at an open configuration.
  • the spacers are fixed on a plate by directly embossing the plate or injection molding of the plate.
  • the materials of the plate and the spacers are selected from polystyrene, PMMA, PC, COC, COP, or another plastic.
  • the inter-spacer spacing in the range of 1 um to 50 um, 50 um to 100 um, 100 um to 200 um or 200 um to 1000 um.
  • the VC sample is an exhaled breath condensate from a human or an animal.
  • the spacers regulating the layer of uniform thickness have a filling factor of at least 1 %, wherein the filling factor is the ratio of the spacer area in contact with the layer of uniform thickness to the total plate area in contact with the layer of uniform thickness.
  • the Young's modulus of the spacers times the filling factor of the spacers is equal or larger than 10 MPa, wherein the filling factor is the ratio of the spacer area in contact with the layer of uniform thickness to the total plate area in contact with the layer of uniform thickness.
  • the thickness of the flexible plate times the Young's modulus of the flexible plate is in the range 60 to 750 GPa-um.
  • ISD4/(hE) the Young's modulus
  • one or both plates comprises a location marker, either on a surface of or inside the plate, that provide information of a location of the plate.
  • one or both plates comprises a scale marker, either on a surface of or inside the plate, that provide information of a lateral dimension of a structure of the sample and/or the plate.
  • one or both plates comprises an imaging marker, either on surface of or inside the plate, that assists an imaging of the sample.
  • the spacers functions as a location marker, a scale marker, an imaging marker, or any combination of thereof.
  • the average thickness of the layer of uniform thickness is about equal to a minimum dimension of an analyte in the sample.
  • the inter-spacer distance is 1 ⁇ or less, 5 ⁇ or less, 7 ⁇ or less, 10 ⁇ 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, 100 ⁇ or less, 200 ⁇ or less, 300 ⁇ or less, 400 ⁇ or less, or in a range between any two of the values.
  • the inter-spacer distance is substantially periodic.
  • the inter-spacer distance is aperiodic.
  • the spacers are pillars with a cross-sectional shape selected from round, polygonal, circular, square, rectangular, oval, elliptical, or any combination of the same.
  • the spacers have are pillar shape and have a substantially flat top surface, wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.
  • each spacer has the ratio of the lateral dimension of the spacer to its height is at least 1.
  • the minimum lateral dimension of spacer is less than or substantially equal to the minimum dimension of an analyte in the sample.
  • the minimum lateral dimension of spacer is in the range of 0.5 um to 100 um.
  • the minimum lateral dimension of spacer is in the range of 0.5 um to 10 um.
  • the spacers have a density of at least 100/mm2. In some embodiments, the spacers have a density of at least 1000/mm2. In some embodiments, at least one of the plates is transparent.
  • At least one of the plates is made from a flexible polymer.
  • the spacers are not compressible and/or, independently, only one of the plates is flexible.
  • the flexible plate for the device of any prior embodiment has a thickness in the range of 10 um to 200 um. In some embodiments, the plate thickness is 0.5 um, 1 um, 5um, 10 um, 25 um, 50 um, 75 um, 100 um, 125 um, 150 um, 175 um, 200 um, 250 um, or a range between any of the two values.
  • the variation in the thickness of the at least a part of the sample in a closed configuration of the plates is less than 30 %, 10 %, 5 %, 3% or 1 %.
  • first and second plates are connected and are configured to be changed from the open configuration to the closed configuration by folding the plates. In some embodiments, the first and second plates are connected by a hinge and are configured to be changed from the open configuration to the closed configuration by folding the plates along the hinge.
  • the first and second plates are connected by a hinge that is a separate material to the plates, and are configured to be changed from the open configuration to the closed configuration by folding the plates along the hinge
  • the first and second plates are made in a single piece of material and are configured to be changed from the open configuration to the closed configuration by folding the plates.
  • the layer of uniform thickness sample is uniform over a lateral area that is at least 100 urn 2 .
  • the layer of uniform thickness sample is uniform over a lateral area that is at least 1 mm 2 .
  • the device is configured to analyze the sample in 60 seconds or less.
  • the final sample thickness device is configured to analyze the sample in 60 seconds or less.
  • the device is configured to analyze the sample in 180 seconds or less.
  • the final sample thickness device is configured to analyze the sample in 180 seconds or less.
  • the device further comprises, on one or both of the plates, one or a plurality of amplification sites that are each capable of amplifying a signal from the analyte or a label of the analyte when the analyte or label is within 500 nm from an amplification site.
  • the final sample thickness device is configured to analyze the sample in 10 seconds or less.
  • the device of any prior embodiment include a step of amplifying the signal from the sample while being in a device that is in a closed configuration of the plates.
  • the signal amplification is achieved without the use of by a physical process and/or biological/chemical amplification.
  • Biological/chemical amplification of the signal include enzymatic amplification of the signal (e.g., used in enzyme-linked immunosorbent assays (ELISAs)) and polymerase chain reaction (PCR) amplification of the signal.
  • the physical signal amplification include plamonics, proximity dependent surface amplification by metal structures or films, and others.
  • the dry binding site comprises a capture agent.
  • the dry binding site comprises an antibody or nucleic acid.
  • the releasable dry reagent is a labeled reagent.
  • the releasable dry reagent is a fluorescently-labeled reagent.
  • the releasable dry reagent is a dye.
  • the releasable dry reagent is a beads.
  • the releasable dry reagent is a quantum dot.
  • the releasable dry reagent is a fluorescently-labeled antibody.
  • the first plate further comprises, on its surface, a first
  • predetermined assay site and a second predetermined assay site, wherein the distance between the edges of the assay site is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the predetermined assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the first plate has, on its surface, at least three analyte assay sites, and the distance between the edges of any two neighboring assay sites is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the first plate has, on its surface, at least two neighboring analyte assay sites that are not separated by a distance that is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the releasable dry reagent is a cell stain.
  • the device further comprises a detector that is an optical detector for detecting an optical signal.
  • the device further comprises a detector that is an electrical detector for detecting an electric signal.
  • the device comprises discrete spacers that are not fixed to any of the plates, wherein at the closed configuration, the discrete spacers are between the inner surfaces of the two plates, and the thickness of the sample is confined by the inner surfaces of the two plates, and regulated by the discrete spacers and the plates.
  • the device further comprises a binding site that has a chemical sensor that is made from a material selected from the group consisting of: silicon nanowire (Si NW); single-walled carbon nanotubes (SWCNT); random networks of carbon nanotubes (RN- CNTs); molecularly capped metal nanoparticles (MCNPs); metal oxide nanoparticles (MONPs); and chemically sensitive field-effect transistors (CHEM-FETs).
  • a chemical sensor that is made from a material selected from the group consisting of: silicon nanowire (Si NW); single-walled carbon nanotubes (SWCNT); random networks of carbon nanotubes (RN- CNTs); molecularly capped metal nanoparticles (MCNPs); metal oxide nanoparticles (MONPs); and chemically sensitive field-effect transistors (CHEM-FETs).
  • a system for rapidly analyzing a vapor condensate sample using a mobile phone comprising:
  • a mobile communication device comprising:
  • system further comprise a light source from either the mobile communication device or an external source.
  • one of the plates has a binding site that binds an analyte, wherein at least part of the uniform sample thickness layer is over the binding site, and is substantially less than the average lateral linear dimension of the binding site.
  • system further comprising:
  • the housing comprises optics for facilitating the imaging and/or signal processing of the sample by the mobile communication device, and a mount configured to hold the optics on the mobile communication device.
  • an element of the optics in the housing is movable relative to the housing.
  • the mobile communication device is configured to communicate test results to a medical professional, a medical facility or an insurance company. In some embodiments, the mobile communication device is further configured to communicate information on the test and the subject with the medical professional, medical facility or insurance company.
  • the mobile communication device is further configured to communicate information of the test to a cloud network, and the cloud network process the information to refine the test results.
  • the mobile communication device is further configured to communicate information of the test and the subject to a cloud network, the cloud network process the information to refine the test results, and the refined test results will send back the subject.
  • the mobile communication device is configured to receive a prescription, diagnosis or a recommendation from a medical professional.
  • the mobile communication device is configured with hardware and software to:
  • c. compare a value obtained from analysis of the test location to a threshold value that characterizes the rapid diagnostic test.
  • At least one of the plates comprises a storage site in which assay reagents are stored.
  • at least one of the cameras reads a signal from the CROF device.
  • the mobile communication device communicates with the remote location via a wifi or cellular network.
  • the mobile communication device is a mobile phone.
  • a method for rapidly analyzing an analyte in a sample using a mobile phone comprising:
  • the analyte comprises a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecule), cells, tissues, viruses, and nanoparticles with different shapes.
  • a molecule e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecule
  • the analyte comprises white blood cell, red blood cell and platelets.
  • the method comprises:
  • the analysis is done by a medical professional at a remote location.
  • the mobile communication device receives a prescription, diagnosis or a recommendation from a medical professional at a remote location.
  • the thickness of the at least a part of VC sample at the closed configuration is larger than the thickness of VC sample deposited on the collection plate at an open configuration.
  • the thickness of the at least a part of VC sample at the closed configuration is less than the thickness of VC sample deposited on the collection plate at an open configuration.
  • the assaying step comprises detecting an analyte in the sample.
  • the analyte is a biomarker.
  • the analyte is a protein, nucleic acid, cell, or metabolite.
  • the assay done in step (b) is a binding assay or a biochemical assay.
  • a method for analyzing an analyte in a vapor condensate sample comprising:
  • the method comprises:
  • each plate has a sample contact surface that is substantially planar, one or both plates are flexible, and one or both of the plates comprise spacers that are fixed with a respective sample contacting surface, and wherein the spacers have:
  • conformable pressing generates a substantially uniform pressure on the plates over the at least part of the sample, and the pressing spreads the at least part of the sample laterally between the sample contact surfaces of the plates, and wherein the closed configuration is a configuration in which the spacing between the plates in the layer of uniform thickness region is regulated by the spacers;
  • the filling factor is the ratio of the spacer contact area to the total plate area; wherein a conformable pressing is a method that makes the pressure applied over an area is substantially constant regardless the shape variation of the outer surfaces of the plates;
  • the method comprises
  • the method comprises removing the external force after the plates are in the closed configuration; and measuring optical signal in the layer of uniform thickness while the plates are the closed configuration.
  • the inter-spacer distance is in the range of 20 urn to 200 urn . In some embodiments, the inter-spacer distance is in the range of 5 urn to 20 urn .
  • a product of the filling factor and the Young's modulus of the spacer is 2 MPa or larger.
  • the surface variation is less than 50 nm.
  • the method further comprising a step of calculating the concentration of an analyte in the relevant volume of sample, wherein the calculation is based on the relevant sample volume defined by the predetermined area of the storage site, the uniform sample thickness at the closed configuration, and the amount of target entity detected.
  • the analyzing step comprise counting the analyte in the sample. In some embodiments, the imaging and counting is done by:
  • the external force is provided by human hand.
  • the method future comprises a dry reagent coated on one or both plates.
  • the layer of uniform thickness sample has a thickness uniformity of up to +1-5%.
  • the spacers are pillars with a cross-sectional shape selected from round, polygonal, circular, square, rectangular, oval, elliptical, or any combination of the same.
  • the spacing between the spacers is approximately the minimum dimension of an analyte.
  • one or both plate sample contact surfaces comprises one or a plurality of amplification sites that are each capable of amplifying a signal from the analyte or a label of the analyte when the analyte or label is within 500 nm from an amplification site.
  • the sample is exhale breath condensate.
  • the sample is a vapor from a biological sample, an environmental sample, a chemical sample, or clinical sample.
  • the analyte comprises a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecules), cells, tissues, viruses, and nanoparticles with different shapes
  • the analyte comprises volatile organic compounds (VOCs).
  • the analyte comprises nitrogen, oxygen, C02, H20, and inert gases.
  • the analyte is stained.
  • the highly uniform thickness has a value equal to or less than 0.5 um. In some embodiments, the highly uniform thickness has a value in the range of 0.5 um to 1 um. In some embodiments, the highly uniform thickness has a value in the range of 1 um to 2 um. In some embodiments, the highly uniform thickness has a value in the range of 2 um to 10 um. In some embodiments, the highly uniform thickness has a value in the range of 10 um to 20 um. In some embodiments, the highly uniform thickness has a value in the range of 20 um to 30 um.
  • it further comprises an enclosure- spacer that encloses a partial or entire VC samples deposited on the collection plate.
  • Embodiment (EBC)-1 SiEBCA (Single-Drop Exhaled Breath Condensate Collector and Analyzer)
  • Exhaled breath condensate (EBC) analysis is a noninvasive method of detecting biomarkers, mainly coming from the lower respiratory tract. EBC is collected during quiet breathing, as a product of cooling and condensation of the exhaled aerosol.
  • An exemplary method of collecting exhaled breath condensate (EBC) using a SiEBCA (Single-drop EBC Collector/Analyzer), as illustrated in Fig. 1 comprises the basic steps:
  • FIG. 1-1 A subject (e.g. human) breathe onto a plate, termed “collection plate”, and the breath condenses into EBC, which are in droplets with different sizes, depending on the breathing time. For a short breathing time most droplets are separated from each other.
  • the surface of the collection plate that collects the EBC is termed the sample surface;
  • the initial droplets are pressed into a thin layer EBC of a thickness that is regulated by the plates and spacers (not shown).
  • SiEBCA can detect and analyze a single drop of EBC deposited on the plate.
  • the substrate plate and “the cover plate” are respectively interchangeable with “the first plate” and “the second plate”.
  • a method of collecting EBC as a basic embodiment of the present invention for collecting EBC from a subject, as illustrated in Fig. 1 , comprises the steps:
  • the exhaled breath condensates on a collection surface of the collection plate to form droplets and/or puddles that have different lateral sizes and different heights, depending upon the surface wetting properties of the collection surface;
  • the open configuration is a configuration in which the two plates are either partially or completely separated apart and the spacing between the plates is not regulated by the spacers;
  • the closed configuration is a configuration, in which: at least a part of the spacers are between the cover plate and the collection plate, and a relevant area of the collection surface of the collection plate is covered by the cove plate and has a plate spacing that is regulated by the spacers;
  • the plate spacing is the spacing between the two inner surfaces (the two surfaces facing each other) of the cover plate and the collection plate, the relevant area is a portion or entire surface of the collection surface, and the collection surface is a portion or entire surface of the collection plate.
  • the EBC sample thickness at the closed configuration is equal to or larger than the EBC sample average thickness at the open configuration.
  • Fig. 4. is an illustration of an embodiment of the devices and the methods of a SiEBCA (Single-drop EBC Collector/Analyzer): (a) having a first plate and a second plate, wherein one or both plate has spacers (shown here: only the first plate has spacers); (b) depositing a sample (only one of many EBC droplets is shown) on the first plate (shown), or the second plate (not shown), or both (not shown) at an open configuration; and (c) (i) using the two plates to spread the sample (the sample flow between the plates) and reduce the sample thickness, and (ii) using the spacers and the plate to regulate the sample thickness at the closed configuration.
  • A2 A device of collecting EBC as a basic embodiment of the present invention for collecting EBC sample from a subject, as illustrated in Fig. 1 , comprises:
  • first plate and a second plate wherein the plates are movable relative to each other into different configurations, and one or both plates are flexible;
  • spacers on one or both of the plates wherein the spacers are fixed with a respective plate, have a predetermined substantially uniform height of 30 urn or less and a predetermined constant inter-spacer distance that is 250 urn or less, and wherein at least one of the spacers is inside the sample contact area;
  • one of the configurations is an open configuration, in which: the two plates are separated apart, the spacing between the plates is not regulated by the spacers, and the EBC sample is deposited on one or both of the plates from a subject;
  • another of the configurations is a closed configuration which is configured after the EBC sample deposition in the open configuration; and in the closed
  • At least part of the EBC sample is compressed by the two plates into a layer of highly uniform thickness, wherein the uniform thickness of the layer is in contact with and confined by the inner surfaces of the two plates and is regulated by the plates and the spacers.
  • the deposition of the EBC sample is by directly exhaling from a subject to one of the plates.
  • the deposition of the EBC sample is by directly exhaling from a subject to both of the plates.
  • Crossing time delay means a time period that it takes from the step (b) EBC deposition of paragraph A1 to the end of the step (c) of paragraph A1 that brings the cover plate and the collection plate to a closed configuration.
  • the covering time delay should be as short as possible to reduce the evaporation of deposited EBC.
  • the covering time delay is equal to or less than 2 sec.
  • the covering time delay is equal to or less than 5 sec.
  • the covering time delay is equal to or less than 10 sec.
  • the covering time delay is equal to or less than 30 sec.
  • the covering time delay is in the range of 30 sec to 300 sec (e.g. 30 to 60 sec, 60 sec to 120 sec, or 120 sec to 300 sec).
  • EBC-1.3 Significant reduction of EBC evaporation rate.
  • One key advantage of the method and device of paragraph A1 and A2 is that, compared to the open configuration of the collection plate and the cover plate, the closed configuraiotn of the plates significantly reduces the surface area of the EBC exposed to the ambient, and hence significantly reduces the EBC sample evaporation rate and significantly increases the time that EBC sample is in liquid form (i.e. the time that EBC sample is not completely evaporated). For example, we have observed that the drying time (the time it takes for the EBC sample to dry out completely) increased from 30 sees at an open configuration to 70 mins, a factor of 140 times longer.
  • guard ring Enclosed Spacers
  • the enclosed spacers or the guard rings can be used to surround the sample to seal off the sample from the ambient.
  • the guard ring can circle an area that is the same as, or larger or smaller than the EBC sample deposited at the open configuration.
  • the guard ring can be configured to further divide an EBC sample into multiple chambers (Fig. 5).
  • Fig. 5 is an illustration of a SiEBCA with both "open spacer" and enclosed spacer, where the open spacer is a post (pillar) while the enclosed spacer is a ring.
  • the enclosed spacer reduces the evaporation of the EBC collected inside the enclosed spacer, since at the closed configuration, the enclosed spacer, the cover plate and the collection plate form an enclosed chamber. If there are only the open spacers but not an enclosed spacer, at the plate closed configuration, the collected EBC still evaporates from the edge of the film formed by the EBC, although such evaporation is much slower than that without a cover plate.
  • the spacers can be an open spacer(s), an enclosed spacer(s), or a combination of thereof.
  • Embodiment EBC-2 EBC Analysis
  • a method of analyzing EBC from a subject for analyzing EBC of a subject comprising:
  • the open configuration is a configuration in which the two plates are either partially or completely separated apart and the spacing between the plates is not regulated by the spacers;
  • the closed configuration is a configuration, in which: at least a part of the spacers are between the cover plate and the collection plate, and a relevant area of the collection surface of the collection plate is covered by the cove plate and has a plate spacing that is regulated by the spacers;
  • the plate spacing is the spacing between the two inner surfaces (the two surfaces face each other) of the cover plate and the collection plate, the relevant area is a portion or entire surface of the collection surface, and the collection surface is a portion or entire surface of the collection plate.
  • the collection plate generally is held at a temperature the same as the ambient, but in some embodiments, the temperatures can be different from the ambient, either higher or lower depending upon the goal of the collection. For example, a temperature lower than the ambient may be used for reducing the EBC evaporation; and a temperature higher than the ambient many bused for evaporating more than that at the ambient temperature is needed.
  • the present invention provides that EBC samples and the target analytes therein can be analyzed by a variety of analysis techniques, depending on the sample volume, species and abundance of the target analyte(s).
  • analysis techniques include spectrometry (e.g., mass spectrometry (MS), ion mobility spectrometry, proton transfer spectrometry, optical absorption spectroscopy systems, and spectrofluorimetry), enzyme based techniques, immunoassay methods (e.g., ELISA, radioimmunoassay, and immune sensors), electronic noses, 2-dimensional protein gel electrophoresis (2D-PEG) followed by chromatographic proteomic microanalysis, nucleic acid (including amplification by PRC (polymerase chain reaction) and isothermal amplification of nucleic acid) by and various types of chemical sensors.
  • spectrometry e.g., mass spectrometry (MS), ion mobility spectrometry, proton transfer spectrometry, optical absorption spectros
  • Another significant advantage of the present invention is that the method and the device of paragraph A1-2, in some embodiments, are used as an EBC analyzer by itself directly or with certain modifications.
  • the EBC analyzer analyzes one or a plurality of target analytes in the EBC.
  • the target analytes are further discussed in Section 3.
  • the modifications made to the method and device of paragraphs A1-A2 include, but not limited to, the following, which can used alone (individually) or in combinations:
  • Binding Sites One or both of the plates have one or plurality of binding site
  • Each (type) of the reagents are in either in well separated locations (the well separation will be defined later).
  • One or both of the plates have one or plurality of binding site
  • binding sites More details of the binding sites, storage sites, amplification sites, and multiplexing sites, as well as their usage for VC and EBC analysis are given in the other part of the disclosure.
  • the modifications made to the method and the device of paragraphs A1-A2 included the use of chemical sensors as binding agents in the binding site for sensing target analytes in the EBC sample, in particular VOC compounds.
  • chemical sensors include, but not limited to. sensors based on conducting polymers and metal oxides, surface acoustic wave sensors, and optical sensors. In some embodiments, they involve the use of
  • the chemical sensors give output in the forms of color change, fluorescence, conductivity, vibration, sound, or any combination thereof, following the interaction of the VOC compound with the sensor.
  • the chemical sensor is made from materials, including, but not limited to, silicon nanowire (Si NW); single-walled carbon nanotubes (SWCNT); random networks of carbon nanotubes (RN-CNTs); molecularly capped metal nanoparticles (MCNPs); metal oxide nanoparticles (MONPs); and chemically sensitive field-effect transistors (CHEM- FETs) that are respectively functionalized for the detection of VOCs or other analytes in the EBC sample.
  • Si NW silicon nanowire
  • SWCNT single-walled carbon nanotubes
  • R-CNTs random networks of carbon nanotubes
  • MCNPs molecularly capped metal nanoparticles
  • MONPs metal oxide nanoparticles
  • CHEM- FETs chemically sensitive field-effect transistors
  • the term “functionalization” refers to the modification to (or the modified properties of) a material that enables it to have specific interaction (e.g., binding, chemical reaction) with certain type(s) of target analytes and bring about a signal indicative of such interaction, thereby to sense the existence and/or abundance of the target analyte.
  • the term “functional group” as used herein refers to the chemical group that is chemically or physically coupled to the material that functionalizes the material.
  • the functional group is made from a protein, an amino acid, a nucleic acid, a lipid, a carbohydrate, a metabolite, any other organic compound, or any other inorganic compound that is able to interact with target analyte in the EBC sample.
  • cross-reactive refers to the fact that a number of reagents/sensors are selectively responsive to a respective range of analytes and the responsive range of each reagent/sensor overlaps partially with at least one of the other reagents/sensors.
  • the device and method of paragraphs A1-A2 include a variety of reagents (sensors), many of which are cross-reactive, and the device and method are utilized in conjunction with a data processing system, including both hardware and software that are capable of pattern recognition.
  • the data processing methods embodied by the data processing system include, but not limited to, principal component analysis, cluster analysis, discriminant function analysis, genetic algorithms, neutral network algorithms, and any other machine learning algorithms.
  • the method and device of prior embodiments are used for collecting EBC samples for analysis by other EBC analyzer(s) according to any of the aforementioned analysis techniques or any combination thereof.
  • a continuous film of the sample is formed after bringing the plates in to the closed configuration.
  • the relevant volume of sample is therefore readily and precisely determined by timing the relevant area by the thickness of the sample film, which is equal to the plate spacing.
  • the plate spacing is equal to or deviating with a small variation from the spacer height.
  • the spacers have a uniform height, and the sample film has a uniform thickness equal to the uniform height.
  • the analyzing step of paragraph AO to A3 comprises: 1) at the closed configuration, taking a first volume of the sample having a first lateral area out of the sample collected in between the cover plate and the collection plate; 2) analyzing the first volume of the sample by an analyzer.
  • the first volume of the sample is equal to the product of the first lateral area and the plate spacing.
  • the first lateral area is pre-determined or pre-known.
  • the device features visual marks that define or border a certain area on the cover plate, the collection plate, or both, within the overlapping area between the two plates, and the first volume of sample is taken according to the marks, e.g., by cutting out along the marks.
  • the sample needs to be a continuous film with no or little empty space within the boundary set by the visual marks.
  • the first lateral area is measured during or after the taking step.
  • the EBC samples collected according to the method and device of paragraphs A1-A2 are analyzed by a combination of both the EBC device itself and other analyzers.
  • Embodiment EBC-3 Exemplary Applications of SiVC/SiEBC
  • Breath tests are among the least invasive methods available for clinical diagnosis, disease state monitoring, health monitoring and environmental exposure assessment.
  • EBC analysis can be used for detection of inflammatory markers, which reflect the state of chronic airways diseases such as chronic obstructive pulmonary disease (COPD), asthma, and cystic fibrosis (CF). EBC analysis can also be used for identification of metabolic, proteomic, and genomic fingerprints of breathing, aiming for an early diagnosis of not only respiratory, but also systemic diseases.
  • COPD chronic obstructive pulmonary disease
  • CF cystic fibrosis
  • a breath matrix from a subject is a mixture of nitrogen, oxygen, C02, H20, and inert gases.
  • the remaining small fraction consists of more than 1000 trace volatile organic compounds (VOCs) with concentrations in the range of parts per million (ppm) to parts per trillion (ppt) by volume.
  • VOCs trace volatile organic compounds
  • these volatile substances may be generated in the body (endogenous) or may be absorbed as contaminants from the environment
  • VOCs in breath varies widely from person to person, both qualitatively and quantitatively.
  • VOCs found to date in human breath are more than 1000, only a few VOCs are common to all humans. These common VOCs, which include isoprene, acetone, ethane, and methanol, are products of core metabolic processes and are very informative for clinical diagnostics.
  • the endogenous compounds found in human breath such as inorganic gases (e.g., NO and CO), VOCs (e.g., isoprene, ethane, pentane, acetone), and other typically nonvolatile substances such as isoprostanes, peroxynitrite, or cytokines, can be measured in breath condensate. Testing for endogenous compounds can provide valuable information concerning a possible disease state. Furthermore, exogenous molecules, particularly halogenated organic compounds, can indicate recent exposure to drugs or environmental pollutants.
  • inorganic gases e.g., NO and CO
  • VOCs e.g., isoprene, ethane, pentane, acetone
  • isoprostanes e.g., peroxynitrite, or cytokines
  • VOCs Volatile Organic Compounds
  • the VOCs that may be assayed as target analytes by the methods and devices provided by the present invention include, but not limited to, biologically generated VOCs (e.g., terpenes, isoprene, methane, green leaf volatiles) and anthropogenic VOCs (e.g., typical solvents used in paints and coatings, like ethyl acetate, glycol ethers, and acetone, vapors from adhesives, paints, adhesive removers, building materials, etc., like methylene chloride, MTBE, and formaldehyde, chlorofurocarbons and perchloroethylene used in dry cleaning, vapor and exhaustive gas from fossil fuels, like benzene and carbon monoxide).
  • biologically generated VOCs e.g., terpenes, isoprene, methane, green leaf volatiles
  • anthropogenic VOCs e.g., typical solvents used in
  • non-volatile compounds have also been lined to or identified as biomarkers of various diseases/conditions.
  • a particular application of the device and method provided by the present disclosure is to assay the glucose level in EBC.
  • Other applications include, but not limited to, detecting the levels of nitrogen reactive species, arachidonic acid metabolites (e.g.,
  • the devices and methods of the present invention also find applications in the detection of drugs of abuse in EBC sample.
  • the drugs of abuse to be detected using the devices and methods of the present invention include, but not limited to, ethanol, cannabis, methadone, amphetamine, methamphetamine, 3,4- methylenedioxymethamphetamine, codeine, 6-acetylmorphine, diazepam, oxazepam, morphine, benzoylecgonine, cocaine, buprenorphine and tetrahydrocannabinol.
  • Table 1 Breath markers in certain diseases or conditions
  • SLE Systemic Lupus Erythematosus
  • CRD Chronic Renal Disease
  • Pentanoic acid 2,2,4-trimethyl-3-carboxyisopropyl, isobutyl ester
  • Certain embodiments of the present invention are related to the applications of the SiEBCA methods and devices for collection and analysis of the vapor condensates other than the EBC.
  • the other moistures include, but not limited to, fog, clouds, steams, etc.
  • the target analysis of these vapor condensates can be for different purpose environmental monitoring, emission control, etc.
  • the sample is a vapor from a biological sample, an environmental sample, a chemical sample, or clinical sample.
  • the devices and methods of the present invention are automatic and high speed, where the steps are performed by machines.
  • the plates are in the form of roll of sheets and are controlled by rollers to put certain area of the plates into an open configuration or a closed configuration.
  • the devices and methods of the present invention are particularly useful for the identification and validation of biomarkers for human
  • the present devices and methods are particularly useful when coupled with data processing system capable of pattern recognition for such purposes.
  • the devices and methods of the present invention are also advantageous to create large sample dataset for refining the algorithms for pattern recognition through machine learning and/or other methodologies.
  • EBC-4 EBC Collection and Analysis without spacers Another aspect of the present invention is to provide devices and methods for collecting and analyzing vapor condensate using the aforementioned collection plate and cover plate but without spacers.
  • the spacers that are used to regulate the sample or a relevant volume of the EBC sample are replaced by (a) positioning sensors that can measure the plate inner spacing, and/or (b) devices that can control the plate positions and move the plates into a desired plate inner spacing based on the information provided the sensors.
  • all the spacers are replaced by translation stage, monitoring sensors and feedback system.
  • the collection plate and the cover plate comprise no spacers at all, and the EBC sample is compressed by the two plates into a thin layer, the thickness of which is regulated by the spacing between the inner surfaces of the plates (the plate spacing).
  • a device for collecting EBC without spacers comprises:
  • the plates are movable relative to each other into different configurations, and one or both plates are flexible;
  • both plates comprise a sample contact area on the respective surface of each plate for contacting EBC sample
  • one of the configurations is an open configuration, in which: the two plates are separated apart, and the EBC sample is deposited on one or both of the plates from a subject;
  • another of the configurations is a closed configuration which is configured after the EBC sample deposition in the open configuration; and in the closed configuration: at least part of the EBC sample is compressed by the two plates into a thin layer, wherein the thin layer is in contact with and confined by the inner surfaces of the two plates.
  • a method of collecting EBC without spacers comprises the steps:
  • step (ii) in the relevant area, a substantial number or all of the droplets or puddles formed in step (b) at the open configuration merge into puddle(s) that (1) have much larger lateral size but in a smaller number than the open configuration and (2) touch both inner surfaces of the cover plate and the collection plate, thereby the thickness of the puddle(s) is confined by the inner surfaces of the plates and equal to the spacing between the inner surfaces, and the total surface area of the deposited EBC exposed to the ambient is significantly reduced;
  • the plate spacing is the spacing between the inner surfaces of the cover plate and the collection plate, the relevant area is a portion or entire surface of the collection surface, and the collection surface is a portion or entire surface of the collection plate.
  • a method of analyzing EBC without spacers comprises the steps:
  • step (i) at least a part of the EBC sample is between the cover plate and the collection plate, and a relevant area of the collection surface of the collection plate is covered by the cove plate; and (ii) in the relevant area, a substantial number or all of the droplets or puddles formed in step (b) at the open configuration merge into puddle(s) that (1) have much larger lateral size but in a smaller number than the open configuration and (2) touch both inner surfaces of the cover plate and the collection plate, thereby the thickness of the puddle(s) is confined by the inner surfaces of the plates and equal to the spacing between the inner surfaces, and the total surface area of the deposited EBC exposed to the ambient is significantly reduced; and
  • the plate spacing is the spacing between the inner surfaces of the cover plate and the collection plate, the relevant area is a portion or entire surface of the collection surface, and the collection surface is a portion or entire surface of the collection plate.
  • the analyzing step (d) of paragraph A6 comprises determining the thickness of the collected EBC sample at the closed configuration after the formation of the thin layer during step (c). In some embodiments, the thickness of the collected EBC sample at the closed configuration is equal to the spacing between the inner surfaces of the two plates.
  • measuring the spacing between the inner surfaces comprises the use of optical interference.
  • the optical interference can use multiple wavelength. For example, the light signal due to the interference of a light reflected at the inner surface of the first plate and the second plate oscillate with the wavelength of the light. From the oscillation, one can determine the spacing between the inner surfaces. To enhance the interference signal, one of the inner surfaces or both can be coated with light reflection material.
  • measuring the spacing between the inner surfaces comprises taking optical imaging (e.g. taking a 2D (two-dimensional)/3D (three-dimensional) image of the sample and the image taking can be multiple times with different viewing angles, different wavelength, different phase, and/or different polarization) and image processing.
  • the analyzing step (d) of paragraph A6 comprises measuring the volume of the collected EBC sample based on the lateral area and the thickness of the thin layer that are determined after the formation of the thin layer during step (c).
  • measuring the entire sample area or volume comprises taking optical imaging (e.g. taking a 2D (two-dimensional)/3D (three-dimensional) image of the sample and the image taking can be multiple times with different viewing angles, different wavelength, different phase, and/or different polarization) and image processing.
  • the sample lateral area means the area in the direction approximately parallel to the first plate and the second plate.
  • the 3D imaging can use the method of fringe projection profilometry (FPP), which is one of the most prevalent methods for acquiring three-dimensional (3D) images of objects.
  • FPP fringe projection profilometry
  • the measuring of the sample area or volume by imaging comprises: (a) calibration of the image scale by using a sample of the known area or volume (e.g., The imager is a smartphone and the dimensions of the image taken by the phone can be calibrated by comparing an image of the a sample of known dimension taken the same phone); (b) comparison of the image with the scale markers (rulers) placed on or near the first plate and second plate (discussed further herein), and (c) a combination of thereof.
  • a sample of the known area or volume e.g., The imager is a smartphone and the dimensions of the image taken by the phone can be calibrated by comparing an image of the a sample of known dimension taken the same phone
  • comparison of the image with the scale markers (rulers) placed on or near the first plate and second plate discussed further herein
  • light may include visible light, ultraviolet light, infrared light, and/or near infrared light.
  • Light may include wavelengths in the range from 20 nm to 20,000 nm.
  • the pressing during step (c) of paragraphs A5-A6 is performed by human hand.
  • the formation and properties of the thin layer is dependent on the pressing force applied during step (c) of paragraphs A5-A6 for bringing the two plates into the closed configuration (as demonstrated in EBC-6).
  • the pressing force applied during step (c) of paragraphs A5-A6 is well adjusted for forming a thin layer of EBC sample between the two plates that has prerequisite parameters.
  • the spacers used on the X-Plate are rectangle pillars in a periodic array with a fixed inner spacer distance (ISD) and uniform spacer height.
  • the pillar spacers have a straight sidewall with a tilt angle from the normal less than 5 degree. Different spacer height, size, inter- spacer distance, shape, and materials are tested.
  • the spacers are fabricated by nanoimprint on a plastic plate, where a mold is pressed directly into the plate.
  • the mold was fabricated by lithography and etching. Examples of the spacers on the plate for SiEBCA.
  • the spacers are fabricated by direct imprinting of the plastic plate surface using a mold, and has a dimension of width, length and height of 30 urn, 40um and 2 urn.
  • EBC Sample deposition All of the EBC samples were deposited on the collection plates by having a human subject directly exhale toward the collection plate which is placed within a few inches away from the subject's mouth.
  • the wetting angles of the different surfaces for different samples were found experimentally as follows: For the liquid of water, PBS, and blood, the contact angle is about 46 degree for untreated glass, 60 degree for untreated PMMA surface, 60 degree for untreated PS (polystyrene) and about 61 degrees for untreated PMMA X- plate. Therefore they are all hydrophilic. But the wetting property of these surfaces can changed to either hydrophilic or hydrophobic by surface treatment. For a good vapor condensate collection, a hydrophilic surface is preferred, which will have, for a given amount of the condensation, smaller surface area by Hand-Press.
  • the present invention has performed various experiments in testing SiVC devices and methods, which are partially described in the PCT [Julia: Inset the info] (ESX-002PCT), which is incorporated herein for its entireity. Some of the experiments are summaried beblow (which used the device with spacers), but the present invention has preformed additional experiments and found that these observations are working for (and can bused) the SiVC device without spacers.
  • Self-Holding means that after a SiEBCA device (plates) is compressed into a closed configuration by an external force (e.g. the force from hand) and after the external force is removed, the SiEBCA device can hold, on its own, the sample thickness unchanged.
  • an external force e.g. the force from hand
  • the SiEBCA device and process can self-hold, as demonstrated in the experiments.
  • Our other experimental test showed that as long as one of the plate is hydrophilic, the SiEBCA plates can self-hold.
  • Drying speed with treated and untreated collection plates was also calculated, which is defined as the retraction length per unit time of the edge of the liquid sample in the X-device at the closed configuration. The calculation shows that with the treated PMMA collection plate, the liquid sample dried at a slower speed (74 um/min) as compared to with the untreated PMMA collection plate (1 17 um/min).
  • the surface treatment of the PMMA plate was performed with oxygen plasma, followed by deposition of 10 nm silicon oxide. In some embodiments, the treatment was performed chemically using trimethoxysilane. AA2.
  • the device of embodiment AAO or AA1 wherein the device further comprises a dry reagent coated on one or both of the plates.
  • the device further comprises, on one or both plates, a dry binding site that has a predetermined area, wherein the dry binding site binds to and immobilizes an analyte in the sample.
  • the device further comprises, on one or both plates, a releasable dry reagent and a release time control material that delays the time that the releasable dry regent is released into the sample.
  • the device further comprises, on one or both plates, one or a plurality of dry binding sites and/or one or a plurality of reagent sites.
  • AA8 The device of any prior embodiment, wherein the sample is a vapor from a biological sample, an environmental sample, a chemical sample, or clinical sample.
  • analyte comprises a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecules), cells, tissues, viruses, and nanoparticles with different shapes.
  • a molecule e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecules
  • cells tissues, viruses, and nanoparticles with different shapes.
  • VOCs volatile organic compounds
  • AA11 The device of any prior embodiment, wherein the analyte comprises nitrogen, oxygen, C02, H20, and inert gases.
  • AA13 The device of any prior embodiment, wherein on one of the sample surface, it further comprises an enclosure-spacer that encloses a partial or entire VC samples deposited on the collection plate.
  • AA15 The device of any prior embodiment, wherein the highly uniform thickness has a value in the range of 0.5 urn to 1 urn.
  • AA16 The device of any prior embodiment, wherein the highly uniform thickness has a value in the range of 1 urn to 2 urn. AA17. The device of any prior embodiment, wherein the highly uniform thickness has a value in the range of 2 urn to 10 urn.
  • AA18 The device of any prior embodiment, wherein the highly uniform thickness has a value in the range of 10 urn to 20 urn.
  • AA20 The device of any prior embodiment, wherein the thickness of the at least a part of VC sample at the closed configuration is larger than the thickness of VC sample deposited on the collection plate at an open configuration.
  • AA21 The device of any prior embodiment, wherein the thickness of the at least a part of VC sample at the closed configuration is less than the thickness of VC sample deposited on the collection plate at an open configuration.
  • AA23 The device of any prior device embodiment, wherein the materials of the plate and the spacers are selected from polystyrene, PMMA, PC, COC, COP, or another plastic.
  • VC sample is an exhaled breath condensate from a human or an animal.
  • the spacers regulating the layer of uniform thickness have a filling factor of at least 1 %, wherein the filling factor is the ratio of the spacer area in contact with the layer of uniform thickness to the total plate area in contact with the layer of uniform thickness.
  • the Young's modulus of the spacers times the filling factor of the spacers is equal to or larger than 10 MPa, wherein the filling factor is the ratio of the spacer area in contact with the layer of uniform thickness to the total plate area in contact with the layer of uniform thickness.. AA29.
  • the thickness of the flexible plate times the Young's modulus of the flexible plate is in the range 60 to 750 GPa-um. AA30.
  • the fourth power of the inter- spacer-distance (ISD) divided by the thickness of the flexible plate (h) and the Young's modulus (E) of the flexible plate, ISD4/(hE), is equal to or less than 106 um3/GPa
  • AA31 The device of any prior paragraph, wherein one or both plates comprises a location marker, either on a surface of or inside the plate, that provides information of a location of the plate.
  • one or both plates comprises a scale marker, either on a surface of or inside the plate, that provides information of a lateral dimension of a structure of the sample and/or the plate.
  • AA33 The device of any prior embodiment, wherein one or both plates comprises an imaging marker, either on surface of or inside the plate, that assists an imaging of the sample.
  • AA34 The device of any prior embodiment, wherein the spacers function as a location marker, a scale marker, an imaging marker, or any combination of thereof.
  • AA35 The device of any prior embodiment, wherein the average thickness of the layer of uniform thickness is about equal to a minimum dimension of an analyte in the sample.
  • the inter-spacer distance is aperiodic.
  • the spacers are pillars with a cross- sectional shape selected from round, polygonal, circular, square, rectangular, oval, elliptical, or any combination of the same.
  • the spacers have a pillar shape and have a substantially flat top surface, wherein, for each spacer, the ratio of the lateral dimension of the spacer to its height is at least 1.
  • each spacer has a ratio of the lateral dimension of the spacer to its height at least 1.
  • AA44 The device of any prior embodiment, wherein the minimum lateral dimension of spacer is less than or substantially equal to the minimum dimension of an analyte in the sample.
  • AA45 The device of any prior embodiment, wherein the minimum lateral dimension of spacer is in the range of 0.5 urn to 100 urn.
  • AA49 The device of any prior embodiment, wherein at least one of the plates is transparent.
  • AA50 The device of any prior embodiment, wherein at least one of the plates is made from a flexible polymer.
  • AA56 The device of any prior embodiment, wherein the first and second plates are connected and are configured to be changed from the open configuration to the closed configuration by folding the plates.
  • AA57 The device of any prior embodiment, wherein the first and second plates are connected by a hinge and are configured to be changed from the open configuration to the closed configuration by folding the plates along the hinge.
  • AA58 The device of any prior embodiment, wherein the first and second plates are connected by a hinge that is a separate material to the plates, and are configured to be changed from the open configuration to the closed configuration by folding the plates along the hinge.
  • AA59 The device of any prior embodiment, wherein the first and second plates are made in a single piece of material and are configured to be changed from the open configuration to the closed configuration by folding the plates.
  • AA60 The device of any prior embodiment, wherein the layer of uniform thickness sample is uniform over a lateral area that is at least 100 urn 2 .
  • AA61 The device of any prior embodiment, wherein the layer of uniform thickness sample is uniform over a lateral area that is at least 1 mm 2 .
  • AA62 The device of any prior embodiment, wherein the device is configured to analyze the sample in 60 seconds or less.
  • the final sample thickness device is configured to analyze the sample in 60 seconds or less.
  • the device further comprises, on one or both of the plates, one or a plurality of amplification sites that are each capable of amplifying a signal from the analyte or a label of the analyte when the analyte or label is within 500 nm from an amplification site.
  • the final sample thickness device is configured to analyze the sample in 10 seconds or less.
  • AA66 The device of any prior embodiment, wherein the dry binding site comprises a capture agent.
  • AA67 The device of any prior embodiment, wherein the dry binding site comprises an antibody or nucleic acid.
  • AA68 The device of any prior embodiment, wherein the releasable dry reagent is a labeled reagent.
  • the first plate further comprises, on its surface, a first predetermined assay site and a second predetermined assay site, wherein the distance between the edges of the assay site is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the predetermined assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the first plate has, on its surface, at least three analyte assay sites, and the distance between the edges of any two neighboring assay sites is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the first plate has, on its surface, at least two neighboring analyte assay sites that are not separated by a distance that is substantially larger than the thickness of the uniform thickness layer when the plates are in the closed position, wherein at least a part of the uniform thickness layer is over the assay sites, and wherein the sample has one or a plurality of analytes that are capable of diffusing in the sample.
  • the releasable dry reagent is a cell stain.
  • the device further comprises a detector that is an optical detector for detecting an optical signal.
  • the device further comprises a detector that is an electrical detector for detecting an electric signal.
  • the device comprises discrete spacers that are not fixed to any of the plates, wherein at the closed configuration, the discrete spacers are between the inner surfaces of the two plates, and the thickness of the sample is confined by the inner surfaces of the two plates, and regulated by the discrete spacers and the plates.
  • the device further comprises a binding site that has a chemical sensor that is made from a material selected from the group consisting of: silicon nanowire (Si NW); single-walled carbon nanotubes (SWCNT); random networks of carbon nanotubes (RN-CNTs); molecularly capped metal nanoparticles (MCNPs); metal oxide nanoparticles (MONPs); and chemically sensitive field-effect transistors (CHEM-FETs).
  • Si NW silicon nanowire
  • SWCNT single-walled carbon nanotubes
  • R-CNTs random networks of carbon nanotubes
  • MCNPs molecularly capped metal nanoparticles
  • MONPs metal oxide nanoparticles
  • CHEM-FETs chemically sensitive field-effect transistors
  • a system for rapidly analyzing a vapor condensation sample using a mobile phone comprising:
  • a mobile communication device comprising: i. one or a plurality of cameras for the detecting and/or imaging the vapor condensate sample;
  • a housing configured to hold the sample and to be mounted to the mobile communication device.
  • the housing comprises optics for facilitating the imaging and/or signal processing of the sample by the mobile communication device, and a mount configured to hold the optics on the mobile communication device.
  • BB7 The system of any prior BB embodiment, wherein the mobile communication device is configured to communicate test results to a medical professional, a medical facility or an insurance company.
  • BB8 The system of any prior BB embodiment, wherein the mobile communication device is further configured to communicate information on the test and the subject with the medical professional, medical facility or insurance company.
  • the mobile communication device is further configured to communicate information of the test to a cloud network, and the cloud network process the information to refine the test results.
  • BB10 The system of any prior BB embodiment, wherein the mobile communication device is further configured to communicate information of the test and the subject to a cloud network, the cloud network process the information to refine the test results, and the refined test results will send back the subject.
  • BB11 The system of any prior BB embodiment, wherein the mobile communication device is configured to receive a prescription, diagnosis or a recommendation from a medical
  • BB14 The system of any prior BB embodiment, at least one of the cameras reads a signal from the CROF device.
  • BB15 The system of any prior BB embodiment, wherein the mobile communication device communicates with the remote location via a wifi or cellular network.
  • BB16 The system of any prior BB embodiment, wherein the mobile communication device is a mobile phone.
  • a method for rapidly analyzing an analyte in a sample using a mobile phone comprising: (a) depositing a sample on the device of any prior BB embodiment;
  • the analyte comprises a molecule (e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecule), cells, tissues, viruses, and nanoparticles with different shapes.
  • a molecule e.g., a protein, peptides, DNA, RNA, nucleic acid, or other molecule
  • analyte comprises white blood cell, red blood cell and platelets.
  • CC6 The method of any prior CC embodiment, wherein the mobile communication device receives a prescription, diagnosis or a recommendation from a medical professional at a remote location.
  • CC8 The method of any prior CC embodiment, wherein the thickness of the at least a part of VC sample at the closed configuration is less than the thickness of VC sample deposited on the collection plate at an open configuration.
  • the assaying step comprises detecting an analyte in the sample.
  • analyte is a protein, nucleic acid, cell, or metabolite.
  • step (b) is a binding assay or a biochemical assay.
  • a method for analyzing an analyte in a vapor condensate sample comprising:
  • each plate has a sample contact surface that is substantially planar, one or both plates are flexible, and one or both of the plates comprise spacers that arefixed with a respective sample contacting surface, and wherein the spacers have: i. a predetermined substantially uniform height,
  • iii a ratio of the width to the height equal or larger than one; iv. a predetermined constant inter-spacer distance that is in the range of 10 ⁇ to 200 ⁇ ;
  • conformable pressing either in parallel or sequentially, an area of at least one of the plates to press the plates together to a closed configuration, wherein the conformable pressing generates a substantially uniform pressure on the plates over the at least part of the sample, and the pressing spreads the at least part of the sample laterally between the sample contact surfaces of the plates, and wherein the closed configuration is a configuration in which the spacing between the plates in the layer of uniform thickness region is regulated by the spacers;
  • the filling factor is the ratio of the spacer contact area to the total plate area; wherein a conformable pressing is a method that makes the pressure applied over an
  • the surface variation is less than 50 nm.
  • any prior DD embodiment further comprising a step of calculating the concentration of an analyte in the relevant volume of sample, wherein the calculation is based on the relevant sample volume defined by the predetermined area of the storage site, the uniform sample thickness at the closed configuration, and the amount of target entity detected.
  • DD10 The method of any prior DD embodiment, wherein the analyzing step comprise counting the ananlyte in the sample.
  • DD11 The method of any prior DD embodiment, wherein the imaging and counting is done by: i. illuminating the cells in the layer of uniform thickness;
  • DD12 The method of any prior DD embodiment, wherein the external force is provided by human hand.
  • DD13 The method of any prior DD embodiment, wherein it future comprises a dry reagent coated on one or both plates.
  • DD15 The method of any prior DD embodiment, wherein the spacers are pillars with a cross- sectional shape selected from round, polygonal, circular, square, rectangular, oval, elliptical, or any combination of the same.
  • one or both plate sample contact surfaces comprises one or a plurality of amplification sites that are each capable of amplifying a signal from the analyte or a label of the analyte when the analyte or label is within 500 nm from an amplification site.
  • any prior CC or DD embodiment wherein the sample is a vapor from a biological sample, an environmental sample, a chemical sample, or clinical sample.
  • the analyte comprises a molecule (e.g. , a protein, peptides, DNA, RNA, nucleic acid, or other molecules), cells, tissues, viruses, and nanoparticles with different shapes.
  • a molecule e.g. , a protein, peptides, DNA, RNA, nucleic acid, or other molecules
  • the present invention includes a variety of embodiments, which can be combined in multiple ways as long as the various components do not contradict one another.
  • the embodiments should be regarded as a single invention file: each filing has other filing as the references and is also referenced in its entirety and for all purpose, rather than as a discrete independent. These embodiments include not only the disclosures in the current file, but also the documents that are herein referenced, incorporated, or to which priority is claimed.
  • CROF Card or card
  • COF Card or card
  • COF Card QMAX-Card
  • Q-Card CROF device
  • COF device COF device
  • QMAX-device CROF plates
  • COF plates COF plates
  • QMAX-plates are interchangeable, except that in some embodiments, the COF card does not comprise spacers; and the terms refer to a device that comprises a first plate and a second plate that are movable relative to each other into different configurations (including an open configuration and a closed configuration), and that comprises spacers (except some embodiments of the COF card) that regulate the spacing between the plates.
  • X-plate refers to one of the two plates in a CROF card, wherein the spacers are fixed to this plate. More descriptions of the COF Card, CROF Card, and X-plate are given in the provisional application serial nos. 62/456065, filed on February 7, 2017, which is incorporated herein in its entirety for all purposes.
  • the devices, systems, and methods herein disclosed can include or use Q-cards, spacers, and uniform sample thickness embodiments for sample detection, analysis, and quantification.
  • the Q-card comprises spacers, which help to render at least part of the sample into a layer of high uniformity.
  • the structure, material, function, variation and dimension of the spacers, as well as the uniformity of the spacers and the sample layer, are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • the devices, systems, and methods herein disclosed can include or use Q-cards for sample detection, analysis, and quantification.
  • the Q-card comprises hinges, notches, recesses, and sliders, which help to facilitate the manipulation of the Q card and the measurement of the samples.
  • the structure, material, function, variation and dimension of the hinges, notches, recesses, and sliders are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • the devices, systems, and methods herein disclosed can include or use Q-cards for sample detection, analysis, and quantification.
  • the Q-cards are used together with sliders that allow the card to be read by a smartphone detection system.
  • the structure, material, function, variation, dimension and connection of the Q-card, the sliders, and the smartphone detection system are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • the sample contact area of one or both of the plates comprises a compressed open flow monitoring surface structures (MSS) that are configured to monitoring how much flow has occurred after COF.
  • MSS comprises, in some embodiments, shallow square array, which will cause friction to the components (e.g. blood cells in a blood) in a sample.
  • the depth of the MSS can be 1/1000, 1/100, 1/100, 1/5, 1/2 of the spacer height or in a range of any two values, and in either protrusion or well form.
  • the devices, systems, and methods herein disclosed can include or be used in various types of detection methods.
  • the detection methods are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • the devices, systems, and methods herein disclosed can be applied to manipulation and detection of various types of analytes (including biomarkers).
  • the analytes and are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • the devices, systems, and methods herein disclosed can be used for various applications (fields and samples).
  • the applications are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes. (9) Cloud
  • the devices, systems, and methods herein disclosed can employ cloud technology for data transfer, storage, and/or analysis.
  • the related cloud technologies are herein disclosed, or listed, described, and summarized in PCT Application (designating U.S.) Nos. PCT/US2016/045437 and PCT/US0216/051775, which were respectively filed on August 10, 2016 and September 14, 2016, US Provisional Application No. 62/456065, which was filed on February 7, 2017, US Provisional Application No. 62/426065, which was filed on February 8, 2017, US Provisional Application No. 62/456504, which was filed on February 8, 2017, all of which applications are incorporated herein in their entireties for all purposes.
  • adapted and “configured” mean that the element, component, or other subject matter is designed and/or intended to perform a given function.
  • the use of the terms “adapted” and “configured” should not be construed to mean that a given element, component, or other subject matter is simply “capable of” performing a given function.
  • subject matter that is recited as being configured to perform a particular function may additionally or alternatively be described as being operative to perform that function.
  • 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 component, feature, detail, structure, embodiment, and/or method is an illustrative, non-exclusive example of components, features, details, structures, embodiments, and/or methods according to the present disclosure.
  • the phrases "at least one of” and “one or more of,” in reference to a list of more than one entity, means any one or more of the entity in the list of entity, and is not limited to at least one of each and every entity specifically listed within the list of entity.
  • “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 the combination of A and B.
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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
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Abstract

La présente invention concerne, entre autres, les procédés, les dispositifs et les systèmes qui peuvent assurer la collecte et l'analyse simple et rapide d'une petite quantité de condensats de vapeur (par exemple, un condensat de souffle exhalé (EBC)).
PCT/US2018/018520 2017-02-16 2018-02-16 Dosage pour condensats de vapeur WO2018152421A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
US16/485,126 US11523752B2 (en) 2017-02-16 2018-02-16 Assay for vapor condensates
US17/980,400 US20230077906A1 (en) 2017-02-16 2022-11-03 Assay for vapor condensates

Applications Claiming Priority (48)

Application Number Priority Date Filing Date Title
US201762459972P 2017-02-16 2017-02-16
US201762460075P 2017-02-16 2017-02-16
US201762460091P 2017-02-16 2017-02-16
US201762460076P 2017-02-16 2017-02-16
US201762459920P 2017-02-16 2017-02-16
US201762460062P 2017-02-16 2017-02-16
US201762460047P 2017-02-16 2017-02-16
US201762460083P 2017-02-16 2017-02-16
US201762460069P 2017-02-16 2017-02-16
US201762460088P 2017-02-16 2017-02-16
US62/460,076 2017-02-16
US62/460,047 2017-02-16
US62/460,091 2017-02-16
US62/459,972 2017-02-16
US62/460,075 2017-02-16
US62/460,083 2017-02-16
US62/459,920 2017-02-16
US62/460,069 2017-02-16
US62/460,062 2017-02-16
US62/460,088 2017-02-16
PCT/US2018/017307 WO2018148342A1 (fr) 2017-02-07 2018-02-07 Dosage et utilisation d'écoulement ouvert comprimé
USPCT/US2018/017307 2018-02-07
PCT/US2018/017492 WO2018148461A1 (fr) 2017-02-09 2018-02-08 Dosage avec amplification
PCT/US2018/017501 WO2018148469A1 (fr) 2017-02-08 2018-02-08 Extraction et dosage de matières bio/chimiques
USPCT/US2018/017494 2018-02-08
USPCT/US2018/017504 2018-02-08
PCT/US2018/017504 WO2018148471A2 (fr) 2017-02-08 2018-02-08 Optique, dispositif et système d'essai
USPCT/US2018/017501 2018-02-08
USPCT/US2018/017489 2018-02-08
USPCT/US2018/017492 2018-02-08
USPCT/US2018/017502 2018-02-08
PCT/US2018/017502 WO2018148470A1 (fr) 2017-02-08 2018-02-08 Collecte et manipulation d'échantillons pour analyse retardée
USPCT/US2018/017499 2018-02-08
PCT/US2018/017494 WO2018148463A1 (fr) 2017-02-08 2018-02-08 Dosage d'hybridation d'acide nucléique
PCT/US2018/017489 WO2018148458A1 (fr) 2017-02-08 2018-02-08 Dosage numérique
USPCT/US2018/017713 2018-02-09
USPCT/US2018/017712 2018-02-09
PCT/US2018/017713 WO2018148607A1 (fr) 2017-02-09 2018-02-09 Dosage utilisant différentes hauteurs d'espacement
USPCT/US2018/017716 2018-02-09
PCT/US2018/017716 WO2018148609A2 (fr) 2017-02-09 2018-02-09 Dosages colorimétriques
PCT/US2018/017712 WO2018148606A1 (fr) 2017-02-09 2018-02-09 Dosage et applications qmax (ii)
USPCT/US2018/018007 2018-02-13
PCT/US2018/018007 WO2018148729A1 (fr) 2017-02-08 2018-02-13 Dispositifs et procédés de dosage à base de carte qmax
PCT/US2018/018108 WO2018148764A1 (fr) 2017-02-08 2018-02-14 Manipulation moléculaire et dosage à température contrôlée
PCT/US2018/017499 WO2018152005A1 (fr) 2017-02-08 2018-02-14 Dosages qmax et applications
USPCT/US2018/018108 2018-02-14
PCT/US2018/018405 WO2018152351A1 (fr) 2017-02-15 2018-02-15 Dosage à changement rapide de température
USPCT/US2018/018405 2018-02-15

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2018/018108 Continuation WO2018148764A1 (fr) 2017-02-08 2018-02-14 Manipulation moléculaire et dosage à température contrôlée

Related Child Applications (2)

Application Number Title Priority Date Filing Date
US16/485,126 A-371-Of-International US11523752B2 (en) 2017-02-16 2018-02-16 Assay for vapor condensates
US17/980,400 Continuation US20230077906A1 (en) 2017-02-16 2022-11-03 Assay for vapor condensates

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WO2018152421A1 true WO2018152421A1 (fr) 2018-08-23

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WO (1) WO2018152421A1 (fr)

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USD898221S1 (en) 2018-11-15 2020-10-06 Essenlix Corporation Assay plate
USD898224S1 (en) 2018-11-15 2020-10-06 Essenlix Corporation Assay plate with sample landing mark
USD898222S1 (en) 2019-01-18 2020-10-06 Essenlix Corporation Assay card
USD898939S1 (en) 2018-11-20 2020-10-13 Essenlix Corporation Assay plate with sample landing mark
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USD910202S1 (en) 2018-11-21 2021-02-09 Essenlix Corporation Assay plate with sample landing mark
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USD898221S1 (en) 2018-11-15 2020-10-06 Essenlix Corporation Assay plate
USD898224S1 (en) 2018-11-15 2020-10-06 Essenlix Corporation Assay plate with sample landing mark
USD897555S1 (en) 2018-11-15 2020-09-29 Essenlix Corporation Assay card
USD898939S1 (en) 2018-11-20 2020-10-13 Essenlix Corporation Assay plate with sample landing mark
USD893469S1 (en) 2018-11-21 2020-08-18 Essenlix Corporation Phone holder
USD910202S1 (en) 2018-11-21 2021-02-09 Essenlix Corporation Assay plate with sample landing mark
USD910203S1 (en) 2018-11-27 2021-02-09 Essenlix Corporation Assay plate with sample landing mark
USD893470S1 (en) 2018-11-28 2020-08-18 Essenlix Corporation Phone holder
USD912842S1 (en) 2018-11-29 2021-03-09 Essenlix Corporation Assay plate
USD898222S1 (en) 2019-01-18 2020-10-06 Essenlix Corporation Assay card
US20210045658A1 (en) * 2019-08-12 2021-02-18 Essenlix Corporation Assay with textured surface
US11712177B2 (en) * 2019-08-12 2023-08-01 Essenlix Corporation Assay with textured surface
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