Devices and methods for sensitive biological sample storage

Abstract

Sample storage devices, kits and methods for storing biological samples are provided. A sample storage device includes an outer housing defining a sample chamber. The sample chamber is configured to contain a biological sample. The sample storage device also includes a deliquescence layer comprising one or more deliquescence materials. The sample storage device also includes a sample membrane, wherein a deliquescent material is associated with the sample membrane to absorb moisture therefrom.

Description

[0001] Cross-references to related applications

[0002] This application is subordinate to and claims priority to U.S. Provisional Patent Application US 63 / 538,628, filed September 15, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention generally relates to apparatus and methods for storing biological samples. Background Technology

[0004] Obtaining biological samples from multiple individuals can often be complicated by the distance between the subject's location and the facility used to analyze the sample. This typically requires the subject to travel to the testing facility to provide the sample for analysis, but this process can be cumbersome depending on the travel distance or the subject's ability to travel. Furthermore, subjects are often located in areas lacking or inconvenient to access testing facilities, and these locations may be in environments relatively unfavorable for sample collection and storage.

[0005] Obtaining samples from subjects at remote locations is often limited by the inability to effectively store them. Transport and environmental conditions can cause samples to degrade before reaching the testing facility. Therefore, there remains a need for alternative devices and methods for storing samples without significant degradation, for example, for subsequent transport to remote testing facilities. Summary of the Invention

[0006] This invention provides apparatus, kits, and methods for collecting and storing biological samples without the need for freezing or refrigeration.

[0007] In one embodiment, the present invention provides a sample storage device comprising an outer shell defining a sample chamber configured to contain a biological sample; a deliquescent layer comprising one or more deliquescent materials; and a sample membrane; wherein the deliquescent material is associated with the sample membrane to be able to absorb moisture from the sample membrane. In some embodiments, the deliquescent material is associated with the sample membrane through direct contact and / or through fluid communication.

[0008] In some embodiments, the housing includes a base and a cover, optionally connected by a hinge and / or a sealing mechanism configured to seal the base and the cover. Optionally, the sealing mechanism is an adhesive tape. In some embodiments, the sample storage device includes a cap and / or a sample collection surface. In some embodiments, the sample collection surface is coupled to the cap.

[0009] Optionally, the deliquescent material is selected from the group consisting of: calcium chloride, magnesium chloride, sodium hydroxide, zinc chloride, ferric chloride, potassium carbonate, potassium phosphate, ferric ammonium citrate, ammonium nitrate, potassium hydroxide, copper sulfate, ammonium persulfate, potassium carbonate, and combinations thereof. In some embodiments, the deliquescent material is calcium chloride. In some embodiments, the sample membrane is a woven material, a spun material, a foam material, a material with multiple capillary channels, or a material having a structure capable of promoting fluid capillary action. Optionally, the sample membrane comprises one or more components of a vitrifying medium, wherein the one or more components are vitrified within the sample membrane. In some embodiments, the sample membrane comprises trehalose, glycerol, or a combination thereof. In some embodiments, the sample storage device comprises a polymer layer comprising a vapor-permeable material.

[0010] In another embodiment, the present invention provides a sampling kit including a sample storage device and a sample collection device. In some embodiments, the sampling kit includes a sample vial containing a vitrified medium.

[0011] In another embodiment, the present invention provides a sample storage method. The method includes inserting a sample into or onto a sample membrane of a sample storage device, and optionally sealing the sample storage device. Optionally, a deliquescent material absorbs moisture from the sample. In some embodiments, the deliquescent material vitrifies the biological sample to produce a storage-stable sample. In some embodiments, the sample is dried to a moisture residual ratio (MRR) of 0.01 to 0.4, optionally about 0.01 to about 0.2, or about 0.09 to about 0.15. In some embodiments, the sample is collected at a location remote from a testing facility. Optionally, the method further includes transporting the sample storage device to a testing facility. In some embodiments, the method further includes restoring the sample to form a restored sample. Optionally, the method further includes testing the restored sample to detect the presence of an analyte, optionally a bioanalyte, optionally cells, bacteria, viruses, yeast, proteins, antibodies, nucleic acids, hormones, fatty acids, steroids, or combinations thereof. In some embodiments, the method further includes quantitatively detecting the presence of the analyte in the sample. Attached Figure Description

[0012] The accompanying drawings are not necessarily drawn to scale; some features may be enlarged or reduced to show details of specific components. Therefore, the specific structural and functional details disclosed herein should not be construed as limiting, but only as a representative basis for teaching those skilled in the art to implement the invention in various ways. Exemplary aspects will be more fully understood from the following detailed description and accompanying drawings, wherein:

[0013] Figure 1A A sample storage device according to one or more embodiments described herein is illustrated schematically.

[0014] Figure 1B A folded configuration is shown according to one or more embodiments described herein. Figure 1A The sample storage device in the middle.

[0015] Figure 1C According to one or more embodiments described herein Figure 1A A cross-sectional view of the device.

[0016] Figure 1D According to one or more embodiments described herein Figure 1B A cross-sectional view of the folded configuration.

[0017] Figure 2A A sample storage device according to one or more embodiments described herein is illustrated schematically.

[0018] Figure 2B According to one or more embodiments described herein Figure 2A A cross-sectional view of the device.

[0019] Figure 2C It includes a sample collection surface according to one or more embodiments described herein. Figure 2A A cross-sectional view of the device.

[0020] Figure 2D According to one or more embodiments described herein Figure 2A A schematic diagram of another sample collection surface in the device.

[0021] Figure 2E The illustrations shown include one or more embodiments according to the present document. Figures 2A to 2D The kit and instructions for use of the sample storage device.

[0022] Figure 3 The illustrations shown include one or more embodiments according to the present document. Figures 1A to 1D The kit and instructions for use of the sample storage device.

[0023] Figure 4 The use of one or more embodiments described herein is illustrated. Figures 1A to 1D Method for sample storage device.

[0024] Figure 5A The concentration of RNA (ng / μL) in cultured cells is shown compared to cells stored with calcium chloride.

[0025] Figure 5B RNA integrity values ​​are shown to indicate a comparison of the quality of RNA isolated from cultured cells with that from cells stored using calcium chloride.

[0026] Figure 5C The percentage of small RNA molecules is shown.

[0027] Figure 5D The percentage of large RNA molecules is shown.

[0028] Figure 5E The results of RNA integrity analysis using gel electrophoresis are shown.

[0029] Figure 6 The percentage of luciferase recovery from freshly collected samples compared to luciferase vitrified onto a sample membrane is shown according to one or more embodiments described herein. Detailed Implementation

[0030] This invention provides a highly advantageous apparatus and method for collecting and optionally storing biological samples, which requires no cold chain storage mechanism, can be deployed in remote locations, is transportable, and / or can be used at locations far from any sample testing facility or apparatus. The provided storage device is configured to maintain and / or enhance the diagnostic value of the sample during transport or storage under varying temperature and humidity conditions. The apparatus and method provided herein utilize a sample membrane, which, through the use of one or more deliquescent materials, enables the sample to be vitrified directly and effectively within or on the sample membrane, thereby forming a storage-stable sample. The device can then be transported to a remote testing facility while maintaining the quality of the acquired sample. These and other advantages of the invention will become more apparent from the following detailed description.

[0031] The following terms or phrases used herein have the following exemplary meanings when relating to at least one embodiment described herein.

[0032] In this article, "vitrification" refers to the process of transforming a material into an amorphous state. The amorphous material may not have any crystalline structure.

[0033] In this document, "vitrification mixture" refers to a heterogeneous mixture of biological material and a vitrification medium containing one or more vitrifying agents and optional other materials.

[0034] In this document, "vitrifying agent" refers to a material that forms an amorphous structure or inhibits crystal formation during the drying and dehydration process of a mixture of vitrifying agent and other materials. One or more vitrifying agents may also provide osmotic protection or otherwise enable cells to survive during dehydration. In some respects, vitrifying agents can be water-soluble solutions capable of producing suitable amorphous structures for the storage / preservation of biological materials. In other respects, vitrifying agents can be absorbed into cells, tissues, or organs.

[0035] In this document, "sample" refers to material that may contain one or more target analytes, target organisms or portions of target organisms, factors or pharmaceutical preparations of any biological origin. The biological sample can be any sample that may carry the target biological factor or target organism. In some embodiments, such as for the storage of biological samples, a volume of biological sample may be placed directly in a sample chamber. The sample can be any sample capable of flowing through the matrix described herein and may carry the target biological factor or target organism. In some embodiments, biological samples include saliva, blood, serum, plasma, urine, bronchoalveolar lavage fluid, sputum, nasal secretions, skin secretions, or combinations thereof. In other embodiments, biological samples are collected from surfaces. In this document, the term "surface" or "environmental surface" generally refers to any surface from which a sample can be collected. Exemplary and non-limiting examples of surfaces include walls, doors, floors, ceilings, drains, appliances, furniture, refrigeration systems, pipes, vents, toilet seats, handles, door handles, handrails, bed rails, countertops, tabletops, dining surfaces, work surfaces, equipment surfaces, clothing, etc.

[0036] Now refer to the attached diagram, Figures 1A to 1D An exemplary embodiment of the sample storage device 100 according to the present invention is illustrated schematically.

[0037] It should be noted that positional terms such as “upper,” “lower,” “proximal,” “farthest,” “top,” and “bottom” are used herein to aid in the description of embodiments of the invention, but are not limiting. The sample storage device 100 may be inverted or positioned differently from the illustrations.

[0038] The sample storage device 100 typically includes a housing 102. The housing 102 is, or substantially, impermeable to water or water vapor. Optionally, the housing 102 is, or substantially, impermeable to air or light. The housing 102 can be formed of any suitable material, including but not limited to polyethylene, polyvinyl chloride, polyvinyl fluoride, polypropylene, polyurethane, fluoropolymers, metals, other impermeable materials, or combinations thereof. The housing 102 optionally defines a sample chamber 104. The sample chamber 104 is optionally sized and shaped to accommodate a sample membrane 106 and one or more additional layers for supporting the storage and / or transport of the sample. Exemplary functions of the additional layers include deliquescence, absorption, support, vapor permeability, and combinations thereof.

[0039] The outer casing 102 optionally includes a base 108 and a cover 110. In some embodiments, the base 108 and the cover 110 are configured to fold together such that the two portions form a bag-like structure surrounding the sample chamber 104, as shown below. Figure 1BAs shown in the diagram. For example, and not bound by theory, the outer shell 102 may be formed with hinges, folding points, or equivalent structures through which the base 108 and the cover 110 can be folded together. Optionally, the base 108 and the cover 110 are configured to seal the sample storage device 100 around the sample chamber 104. Figure 1B As shown, the sealing mechanism 112 may be disposed on the base 108 and / or cover 110 surrounding the sample chamber 104. Any suitable sealing mechanism may be employed. An exemplary seal may be formed from tape including heat-activated adhesives, pressure-sensitive adhesives, gaskets, etc.

[0040] The sample storage device 100 may include a sample membrane 106. The sample membrane 106 may optionally be attached to the sample storage device 100. Alternatively, the sample membrane 106 may be a separate component configured to be inserted into the sample storage device 100. In such an embodiment, the sample membrane 106 may be included within a kit 300, which will be described in further detail below.

[0041] The sample membrane 106 may be a membrane, a scaffold, multiple capillary channels, a woven material, a spun material, a foam material, or any other structure operable to contain or support a biological sample for vitrification. Optionally, the sample membrane 106 may be in the form of a random or ordered fibrous network structure that defines channels or other continuous or discontinuous pathways for the entry of fluid or biological samples. Optionally, the sample membrane 106 may be in the form of a nonwoven fibrous web, optionally having a high porosity (e.g., greater than 50% by volume). In some embodiments, the sample membrane 106 is a foam material, such as polyethersulfone (PES), polyurethane (PU), polyethylene (PE), polypropylene (PP), polystyrene, etc. Illustrative examples of suitable bases can be found in U.S. Patent Application Publication No. 2023 / 0082652, International Patent Application Publication No. WO 2020 / 086812, International Patent Application No. PCT / US2022 / 035892 and International Patent Application No. PCT / US2021 / 060164, which also teach methods for vitrifying molecules or incorporating molecules into a matrix or substrate, the entire contents of which are incorporated herein by reference.

[0042] In some embodiments, the sample membrane 106 is formed of one or more materials, and one or more vitrification components may be incorporated into or onto said material at one or more locations. Exemplary examples of materials used for the sample membrane 106 include, but are not limited to: collagen, elastin, hyaluronic acid and its derivatives, sodium alginate and its derivatives, chitosan and its derivatives, gelatin, starch, cellulose polymers (e.g., nitrocellulose (e.g., Sartorius CN95), methylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, carboxymethylcellulose, cellulose acetate phthalate, cellulose acetate succinate, hydroxypropyl methylcellulose phthalate), and poly(citrate glycol). (e.g., poly(octyl glycol citrate),) casein, dextran and its derivatives, poly(caprolactone), poly(hydroxy acids), poly(L-lactide), poly(D,L-lactide), poly(D,L-lactide-co-glycolic acid), poly(L-lactide-co-glycolic acid), copolymers of lactic acid and glycolic acid, copolymers of ε-caprolactone and lactide, copolymers of glycolide and ε-caprolactone, copolymers of lactide and 1,4-dioxane-hexanone, containing monomers D-lactide, L-lactide, D,L- Polymers and copolymers of one or more residue units of lactide, glycolide, ε-lactide, methyl methacrylate, 1,4-dioxane or 1,5-dioxane-heptanone; polyglycolic acid, polyhydroxybutyrate, polyalkyl carbonate and polyorthoester; polyester; poly(hydroxyvalerate); poly(p-dioxane); poly(ethylene terephthalate); poly(malic acid); poly(tartaric acid); polyanhydride; polyphosphazene; poly(amino acid); and copolymers of the above polymers, as well as blends and combinations thereof. Illustratively, such polymers can be seen in the relevant descriptions in U.S. Patent Application Publication No. 2018 / 0125990 and its cited references, including Polymers in Controlled Drug Delivery (Illum, L. and Davids, SS, eds., Wright Publishing, Bristol, 1987); Arshady, J. Controlled Release, 17:1-22 (1991); Pitt, Int. J. Phar., 59:173-96 (1990); Holland et al., J. Controlled Release, 4:155-80 (1986).

[0043] The material used for the sample membrane 106 herein may optionally be formed into a porous network, such as a filter membrane, a symmetrical network, or other such porous sheet-like material. Illustratively, the sample membrane 106 material is formed as a fiber network. This fiber network can be formed using any suitable method. For example, without limitation, in some embodiments, the sample membrane 106 is formed by electrospinning. During electrospinning, the desired material is placed in a desired solvent (e.g., 2,2,2-trifluoroethanol (TFE) or hexafluoroisopropanol (HFIP)) and electrospinned to form fibers having the desired cross-sectional dimensions and lengths, and arranged in a desired orientation (optionally random) to obtain the desired pore size (average distance between strands) to allow materials, analytes, active agents, etc., to pass through the network or be retained within or trapped by the network. Optionally, the sample membrane 106 comprises two or more layers of material, optionally three, four, five, six, or more layers of material.

[0044] In some embodiments, the sample membrane 106 defines one or more pores. Optionally, the pore size is adapted to control sample flow or regulate the size of molecules that can pass through the sample membrane 106. It should be understood that in some embodiments, the pore size can be adjusted based on the desired molecular size or sample flow. In some aspects, the pore size within the sample membrane 106 ranges from about 5 micrometers (μm) to about 500 μm, or any value or range therebetween. In some aspects, the average opening size of the pores can be from about 20 μm to about 0.1 μm, including about 19 μm, 18 μm, 17 μm, 16 μm, 15 μm, 14 μm, 13 μm, 12 μm, 11 μm, 10 μm, 9 μm, 8 μm, 7 μm, 6 μm, 5 μm, 4 μm, 3 μm, 2 μm, 1.0 μm, 0.9 μm, 0.8 μm, 0.7 μm, 0.6 μm, 0.5 μm, 0.4 μm, 0.3 μm, and 0.2 μm. In some aspects, the pore size of the sample membrane 106 is selected to allow the passage of any markers or target analytes present in the liquid and / or sample. In multilayer embodiments, different layers of the sample membrane 106 can define pores with different pore sizes.

[0045] In some embodiments, the sample membrane 106 defines capillary channels. The capillary channels may have a length optionally defined by the thickness of the sample membrane 106 forming the channels or by one or more individual channels themselves. The capillary channel length may optionally be about one millimeter or less, but should not be construed as limited to such a size. Optionally, the capillary channel length is about 0.1 μm to about 1000 μm, or any value or range therebetween. Optionally, the capillary channel length is about 5 μm to about 100 μm, optionally about 1 μm to about 200 μm, and / or optionally about 1 μm to about 100 μm. The capillary channel length may optionally be approximately 5 μm, 10 μm, 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm, 50 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, 80 μm, 85 μm, 90 μm, 95 μm, or 100 μm. In some aspects, the capillary channel lengths in the multiple capillary channels may differ from each other and may optionally exhibit non-uniform variation.

[0046] The cross-sectional area of ​​capillary channels can be approximately 2000 μm. 2 Or smaller. Optionally, the cross-sectional area is approximately 0.01 μm. 2 Approximately 2000μm 2 Optionally, it can be approximately 100 μm. 2 Approximately 2000μm 2 , or any value or range thereof. Optionally, the cross-sectional area of ​​the capillary channel is approximately 100 μm. 2 200μm 2 300μm 2 400μm 2 500μm 2 600μm 2 700μm 2 800μm 2 900μm 2 1000μm 2 1100μm 2 1200μm 2 1300μm 2 1400μm 2 1500μm 2 1600μm 2 1700μm 2 1800μm 2 1900μm 2 or 2000μm 2 Or smaller.

[0047] In some aspects, the sample membrane 106 is a hydrophilic material or includes a hydrophilic material. In other aspects, the sample membrane 106 is a hydrophobic material or includes a hydrophobic material. In some embodiments, the sample membrane 106 may be treated to have hydrophilicity or stronger hydrophilicity, such as by exposure to plasma.

[0048] In some embodiments, the sample membrane 106 is or includes polycaprolactone (PCL), collagen, or a combination thereof. It should be understood that such water-stable polymers are capable of forming networks or fibers, allowing one or more components to be vitrified thereon or therein. Therefore, in some embodiments, the sample membrane 106 possesses sufficient stability in an aqueous environment to serve as a surface suitable for vitrifying an aqueous vitrification medium containing one or more target molecules, such as sample factors and / or detection factors.

[0049] In some embodiments, for example Figure 1C As shown, the sample chamber 104 is divided into one or more additional layers. Each of these additional layers can be in direct contact with another layer and / or can be separated from other layers by an air gap. Optionally, the sample storage device 100 includes a polymer layer 114 for supporting the sample membrane 106. Optionally, the polymer layer 114 is formed of a vapor-permeable material, such as spunbond polyethylene polymer sheet (i.e., TYVEK®). Other exemplary and non-limiting vapor-permeable materials include expanded polytetrafluoroethylene, microporous polypropylene membranes, hydrophilic polyether-ester block copolymers (e.g., SYMPATEX®), spunbond polypropylene, combinations of spunbond polypropylene and meltblown polypropylene, and combinations thereof, but any other suitable vapor-permeable material is contemplated and may be used.

[0050] Sample storage device 100 typically includes a deliquescent layer 116. In some embodiments, the deliquescent layer 116 is in fluid communication with sample membrane 106, and optionally in vapor communication. In some embodiments, the deliquescent layer 116 is in direct contact with sample membrane 106. In some embodiments, the deliquescent layer 116 and sample membrane 106 are separated by a gap, optionally configured to allow fluid (optionally via vapor) contact between the sample membrane and the deliquescent layer 116.

[0051] In some embodiments, vitrification is achieved by removing moisture from the sample membrane 106 to the deliquescent material until the sample enters a glassy state, as described in more detail herein. Optionally, the deliquescent layer 116 aids in vitrification of the sample by drawing water through the polymer layer 114 and absorbing water. In other embodiments, vitrification of the sample is achieved by alternative methods, such as those disclosed in U.S. Patent Application Publication No. 2023 / 0082652, International Patent Application Publication No. WO 2020 / 086812, International Patent Application Publication No. PCT / US2022 / 035892, and International Patent Application Publication No. PCT / US2021 / 060164. In other embodiments, instead of achieving complete vitrification of the sample, sufficient degree of vitrification is achieved to prevent degradation for a time period at least equal to or greater than any desired storage time, optionally from 1 to 10 days, optionally less than 10 days, optionally 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 days.

[0052] The deliquescent layer 116 typically comprises one or more deliquescent materials. In this document, “deliquescence” or “hygroscopicity” refers to the phenomenon where a substance absorbs moisture from its surrounding environment, where, in the presence of sufficient moisture, the substance undergoes a phase transition from a solid or crystalline material to a liquid. In some respects, this phase transition occurs irreversibly (at standard temperatures and pressures), thereby trapping the fluid and preventing its re-emission. However, it should be understood that the deliquescent material does not need to undergo a complete phase transition to facilitate adequate vitrification of the sample. For example, the amount of deliquescent material may optionally be sufficient, or other mechanisms may be provided, such that the amount of fluid removed from the sample and entering the deliquescent material is sufficient to completely or partially vitrify the sample, but not necessarily to cause a complete phase transition of the entire deliquescent material. However, the deliquescent material may optionally trap water from the sample and prevent the re-transfer of such water back to the sample, thereby maintaining the achieved level of vitrification in the sample and promoting stable storage of the vitrified sample in the apparatus until the desired time, such as the time for analysis.

[0053] Exemplary and non-limiting deliquescent materials that can be used in the apparatus and methods provided herein include, but are not limited to, calcium chloride, magnesium chloride, sodium hydroxide, zinc chloride, ferric chloride, potassium carbonate, potassium phosphate, ferric ammonium citrate, ammonium nitrate, potassium hydroxide, copper sulfate, ammonium persulfate, potassium carbonate, and combinations thereof. In some embodiments, the deliquescent material is calcium chloride.

[0054] Many sample types, including but not limited to biological and pharmaceutical samples, require protection from moisture to prevent degradation, extend shelf life, and prevent bacterial and fungal growth. Deliquescent materials effectively absorb moisture. Such materials can absorb several times their own weight in water, and in the presence of sufficient moisture, if the air humidity and temperature are sufficiently high, these materials will dissolve to form a liquid brine. The liquid brine can optionally be contained within a collector or mixed with modified starch to form a gel within its packaging. Furthermore, deliquescent materials are self-limiting, allowing them to provide continuous protection without depletion during periods of high humidity. Additionally, deliquescent materials are environmentally safe and can be handled without hazardous materials disposal procedures. Deliquescent materials can optionally absorb larger amounts of moisture when the relative humidity of the surrounding air is high. For example, without any theoretical limitations, at 50% relative humidity, calcium chloride absorbs 150% of its own weight in moisture. As relative humidity increases, its moisture absorption also increases. For example, at 85% relative humidity, calcium chloride can absorb up to 600% of its own weight in moisture.

[0055] The device described herein also prevents the absorbed moisture from evaporating again. See still for details. Figure 1C The sample storage device 100 optionally includes an absorbent layer 118. As the deliquescent material absorbs moisture, it forms a liquid solution. The absorbent layer 118 can trap this liquid solution, thereby further preventing moisture re-evaporation and potential damage to the sample. In some embodiments, the absorbent layer 118 includes a desiccant, hydrogel, sponge, nonwoven material, cotton, superabsorbent polymer, or similar material. In some embodiments, the absorbent layer 118 includes a desiccant, such as silica gel, clay desiccant, or similar material.

[0056] In some embodiments, the base 108 and the cover 110 each include a plurality of additional layers. Optionally, the base 108 and the cover 110 each include a polymer layer 114, a deliquescent layer 116, and an absorbent layer 118.

[0057] Now for reference Figures 2A to 2D Another embodiment of the sample storage device 200 is schematically illustrated. In this embodiment, the sample storage device 200 includes an outer housing 202 defining a sample chamber 204. The sample storage device 200 also includes a cap 208. The sample storage device 200 includes a sample collection surface 206. The sample collection surface 206 may optionally be formed of the same material as described above with respect to sample membrane 106.

[0058] The sample collection surface 206 is optionally associated with an elongated rod 254. The elongated rod 254 can be made of a variety of materials, such as plastic, metal, wood, or combinations thereof. In some embodiments, the elongated rod 254 is flexible, allowing it to reach a sampling site that would otherwise be inaccessible. In other embodiments, the elongated rod 254 can be relatively rigid, allowing force to be applied to the elongated rod 254 to collect a sample.

[0059] The sample collection surface 206 may optionally be in the form of a swab or a device for contacting the sampling site to collect a sample. Generally, the collection surface 206 is adapted to receive a sample. In embodiments, the collection surface 206 is formed of one or more liquid-permeable materials, such as cotton, fibers, sponges, foam materials, nonwoven fabrics, bamboo, hydrophilic or hydrophobic polymers, and similar materials. In embodiments, the collection surface 206 is sterile to prevent sample contamination or inaccurate results.

[0060] The collecting surface 206 can be attached to the elongated rod 254 using any suitable mechanism, including but not limited to fasteners, adhesives, heat seals, thermal bonding, etc. In some embodiments, the collecting surface 206 is attached to the elongated rod 254 by entanglement of a liquid-permeable material on and / or around the elongated rod 254. In some embodiments, the collecting surface 206 is attached to the elongated rod 254 by one or more threaded fasteners.

[0061] The size and shape of the sample acquisition surface 206 can be designed to be fully or partially placed in the sample chamber 204 after the sample is acquired. In some embodiments, for example... Figure 2D As shown, the sample collection surface 206 is attached to the cap 208. In such an embodiment, after sample collection, the swab is tightened or otherwise inserted back into the tube for storage. In other embodiments, for example... Figure 2C As shown, the sample acquisition surface 206 is spaced apart from the sample storage device 200. For example, in some embodiments, the sample acquisition surface 206 is part of the sample acquisition device 252, which is provided as part of a kit, as described in more detail herein.

[0062] In some embodiments, the collection surface 206 may be a sample membrane 106 or may be used as or coupled to a sample membrane, allowing fluid to be transferred from the collection surface 206 to the sample membrane 106. Optionally, the sample membrane 106 is coupled to the elongated rod 254. In some embodiments, the sample membrane 106 is disposed on the proximal surface of the collection surface 206. In some embodiments, the sample membrane 106 is at least partially disposed between the elongated rod 254 and the collection surface 206. Optionally, the sample membrane 106 is a layer disposed beneath the collection surface 206. In some embodiments, the collection surface 206 partially or completely covers the sample membrane 106.

[0063] Still referencing Figure 2A-2D The sample storage device 200 typically includes a polymer layer 214 that separates the sample chamber 204 from the housing 202 and the deliquescent layer 216. Optionally, the polymer layer 214 is formed of a vapor-permeable material, such as spunbond polyolefin polymer sheets, illustratively but not limited to those sold under TYVEK®.

[0064] Sample storage device 200 typically includes one or more deliquescent layers 216. The deliquescent layer 216 contains one or more deliquescent materials. The deliquescent material in the deliquescent layer 216 may be formed of the same material described above with respect to deliquescent layer 116. In some embodiments, the deliquescent material in the deliquescent layer 216 is calcium chloride.

[0065] In some embodiments, the sample storage device 200 is provided as part of the kit 250. For example... Figure 2E As shown, kit 250 typically includes sample storage device 200 and sample collection device 252.

[0066] The sample collection device 252 typically includes an elongated rod 254 and a sample collection surface 206 coupled to the elongated rod 254. Generally, the collection surface 206 is adapted to receive biological samples. In some embodiments, the collection surface 206 is formed of one or more liquid-permeable materials, such as cotton, fibers, sponges, foam materials, nonwoven fabrics, bamboo, hydrophilic polymers, and similar materials. Optionally, the collection surface 206 is sterile to prevent sample contamination or inaccurate results.

[0067] During operation, the sample acquisition device 252 is used to contact the sample. The sample is acquired by bringing the acquisition surface 206 into contact with the sampling site. After sample acquisition, the sample acquisition device 252 can be inserted into the tube. The elongated rod 254 can optionally be broken off to ensure that the sample acquisition device 252 can be placed into the sample storage device 200. The cap 208 can then be installed to seal and isolate the sample chamber 204 from the external environment and promote vitrification of the sample within or on the sample membrane 106 through the action of the deliquescent material within the device.

[0068] In some embodiments, the sample storage device 100 is provided as part of the kit 300. For example... Figure 3 As shown, kit 300 typically includes a sample storage device 100 and a sample acquisition device 302. It should be understood that the reference numerals for the various components are... Figure 3 The reference numerals are labeled in the first frame; however, for clarity and to avoid repetition, the reference numerals are omitted in subsequent frames, as the components can be visually identified from the context of the figures.

[0069] The sample collection device 302 typically includes an elongated rod 304 and a sample collection surface 306 coupled to the elongated rod 304. Generally, the collection surface 306 is adapted to receive a sample. Optionally, the collection surface 306 is formed of one or more liquid-permeable materials, such as cotton, fibers, sponges, foam materials, nonwoven fabrics, bamboo, hydrophilic polymers, etc. In embodiments, the collection surface 306 is sterile to prevent sample contamination or inaccurate results.

[0070] During operation, the sample collection device 302 is used to contact the sample. Collection can be performed by bringing the collection surface 306 into contact with the sample, optionally by wiping the subject or a portion thereof, dripping sample material onto the collection device or other apparatus, or by other methods. For example, the collection surface 306 can be gently wiped or rotated on the sampling site to allow it to absorb or collect the sample. Exemplary sampling sites include, but are not limited to, skin, nails, mucous membranes, wounds, and environmental surfaces.

[0071] In some embodiments, the sample is mixed with the vitrification medium before or during contact with the sample membrane 106. In some embodiments, for example... Figure 3 As shown, kit 300 includes one or more vials 308 containing a vitrification medium. The vitrification medium may be provided in liquid or dry form. In some embodiments, the sample is mixed with the vitrification medium to form a vitrification solution. If the biological sample is a liquid biological sample, the vitrification medium may be dissolved in the liquid sample. In short, the vitrification medium comprising one or more sample factors to be vitrified is combined with a matrix such as sample membrane 106 and placed or applied onto said sample membrane 106.

[0072] A vitrifying medium may include at least one vitrifying agent. Illustrative examples of vitrifying agents include, but are not limited to, dimethyl sulfoxide, glycerol, sugars (e.g., trehalose), polyols, methylamines, betaines, antifreeze proteins, synthetic anti-nucleating agents, polyvinyl alcohol, cyclohexanetriol, cyclohexanediol, inorganic salts, organic salts, ionic liquids, or combinations thereof. In some aspects, one, two, three, four, or more vitrifying agents are included in the vitrifying medium.

[0073] Vitrifying agents are contained in the vitrification medium at concentrations that depend on their properties. In some respects, the concentration of the vitrifying agent is below a concentration that would be toxic to the vitrified biological sample. As used herein, "toxic" means that the sample will not achieve functional or biological activity after subsequent use, or that the biological sample is unsuitable for subsequent analysis. In several respects, the concentration of the vitrifying agent is greater than or equal to 500 micromoles (μM) and less than or equal to 6 moles (M), or any value or range therebetween. As an example, the concentration of trehalose is greater than or equal to 1 millimole (mM) and less than or equal to 6 M, optionally greater than or equal to 150 mM and less than or equal to 6 M. In some respects, the total concentration of all vitrifying agents combined is greater than or equal to 1 mM and less than or equal to 6 M, optionally greater than or equal to 1 mM and less than or equal to 6 M.

[0074] In some aspects, it is conceivable that the vitrification medium may further include other components, such as, by way of example but not limited to, water or other solvents, buffers, one or more salts, RNase or DNase inhibitors, or combinations thereof. Buffers are any reagents with a pKa of 6 to 8.5 at 25°C. Illustrative examples of buffers include choline, betaine, HEPES, TRIS, PIPES, MOPS, etc. In some aspects, buffers are buffers containing large organic ions (greater than 120 kDa), such as choline, betaine, or HEPES. In embodiments including buffers, the buffers are provided at a concentration suitable for stabilizing the pH of the vitrification medium at a desired level.

[0075] The salt may include, for example, but not limited to, magnesium salts, sodium salts, potassium salts, chloride salts, or combinations thereof. When included in a vitrification medium, the salt may be provided at a concentration greater than or equal to 1 millimole (mM) and less than or equal to 500 mM.

[0076] The vitrification medium may optionally include sugars (e.g., trehalose) and may also include a combination of divalent metal ions and chelating agents. Illustrative examples of divalent metal ions include salts of Ca, Mg, Co, Fe, Zn, Mn, etc. Exemplary salts include chlorides, sulfates, acetates, etc. Chelating agents include polyols, ethylenediaminetetraacetic acid (EDTA), ethylene glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA), etc. In the vitrification solution, the molar ratio of the divalent metal salt to the chelating agent may be from 10:90 to 90:10, optionally 50:50.

[0077] Materials used in vitrifying media, detection agents, indicators, buffers, or any other desired materials may optionally be vitrified into or onto a particular device or portion thereof, optionally onto a collection surface, sample membrane, or other portion, optionally in accordance with the teachings of U.S. Patent 10,433,540. Upon contact with a fluid sample, the fluid in the sample causes the vitrifying agent to re-dissolve / restore to form a vitrified solution. The vitrified solution is then placed in the device provided herein, wherein the vitrified solution is subsequently partially or completely vitrified by the action of a deliquescent material.

[0078] The sample, alone or in combination with a vitrification solution (the vitrification medium), is applied to the sample membrane 106. The sample storage device 100 is then sealed to form a seal and stored or transported prior to analysis.

[0079] In other embodiments, the sample storage device 100 receives the sample via a fluid connection. For example... Figure 4 As shown, in some embodiments, biological samples are collected from a patient. For example, and without being theoretically constrained, the sample can be collected into a collection tube 402. The collection tube 402 is then fluidly coupled to a sample storage device 100. The sample can then be transferred to and sealed in the sample storage device 100. Optionally, the sample storage device 100 is under vacuum, such that when the sample storage device 100 is coupled to the collection tube 402, the sample is drawn into the sample storage device and comes into contact with the sample membrane. In other aspects, the sample storage device 100 is used to directly collect samples by means of vacuum action to transfer samples from a subject or surface into the sample storage device 100. It should be understood that... Figure 4 The sample storage device is illustrated as configured in Figure 1, but this is not limiting. The sample storage device may optionally be similar to... Figure 2B The sample membrane 106 is in the form of a tube and can be connected to a system or device that subsequently comes into contact with the patient. The sample membrane 106 can be laminated onto the polymer layer 214 such that when a sample enters the sample storage device, the sample contacts the sample membrane and is then vitrified for final storage and subsequent use.

[0080] Sample storage devices 100 and 200 are optionally used to store samples or portions thereof in a fully or partially vitrified form, such that the samples remain sufficiently stable for a required period of time prior to analyte detection. This document also provides methods for detecting one or more sample factors in a sample. These methods include, but are not limited to, inserting a sample into or onto a sample membrane of the device or kit described herein, and optionally sealing the sample storage device. Optionally, the sample storage device is penetrated by a collection device, such that a sample is transferred into the sample storage device to contact a sample membrane as provided herein. The sample storage device, comprising a deliquescent material, then absorbs moisture from the sample, such that, in the presence of one or more vitrifying agents, the sample analyte is subsequently partially or fully vitrified. The deliquescent material vitrifies the sample to produce a storage-stable sample. A storage-stable sample, as defined herein, is a sample that remains (physically or functionally) no more than 10% of its volume during storage, optionally from 1 to 10 days, or as described elsewhere herein.

[0081] One method involves generating a storage-stable sample with residual moisture content. A storage-stable sample is defined as one in which the functional degradation of the analyte does not exceed 10% of its activity over a period not exceeding 10 days. Activity is based on the analyte's expected activity. For example, the activity of an antibody can be defined as its ability to bind to a target; the activity of an enzyme can be defined as its ability to act on a substrate.

[0082] The residual moisture content can optionally be quantified as the residual moisture ratio (MRR), which is defined as: the actual weight of the dried film after vitrification (x... a Subtract the amount including the analyte (x) t The weight of the dry components (e.g., trehalose, glycerol, BSA) of the glassization buffer is divided by x. a Therefore, MRR = (x a − x t ) / x a MRR is a ratio and has no unit. The MRR of the material may optionally be from 0.001 to 0.4, or any value or range therebetween. Optionally, the MRR is from 0.005 to 0.2. Optionally, the MRR is from 0.01 to 0.08. Optionally, the MRR is from 0.01 to 0.15. Optionally, the MRR is from 0.08 to 0.15. Optionally, the MRR is from 0.09 to 0.2. Optionally, the MRR is from 0.09 to 0.15.

[0083] Optionally, one method includes collecting samples at a remote location. A remote location refers to a location where the appropriate reagents or equipment for analyzing the samples are not available, and therefore the sample storage device needs to be transported from a remote location to the testing facility for subsequent sample analysis.

[0084] At the testing facility, the methods provided herein may optionally include subjecting a sample to one or more analytical methods to detect and / or quantify the presence of one or more analytes in the sample. One method may optionally include reconstituted / redissolved the sample by contacting the stored, stable sample with one or more eluents. The eluent may optionally be any liquid suitable for dissolving vitrified materials in a matrix. Optionally, the eluent is water or an aqueous material. Optionally, the eluent is or includes one or more organic solvents. Optionally, the eluent includes one or more buffers.

[0085] Illustrative examples of elution buffers include, but are not limited to, choline, betaine, HEPES, TRIS, PIPES, MOPS, etc. Optionally, the elution buffer may include, for example, but not limited to, magnesium salts, sodium salts, potassium salts, chloride salts, or combinations thereof. When included in the elution buffer, the salt may be provided at a concentration greater than or equal to 1 mmol (mM) to less than or equal to 500 mM. Optionally, the components of the elution buffer may be provided at concentrations suitable for stabilizing the pH of the elution buffer at a desired level.

[0086] The reconstituted sample may optionally be analyzed using one or more analytical techniques. Analytical techniques may include, but are not limited to, luminescence detection, mass spectrometry, PCR, Western blotting, DNA blotting, flow cytometry, enzymatic detection, spectrometry, or other techniques, or combinations thereof.

[0087] Optionally, the detection of the analyte relies on an enzyme / substrate reaction. In an embodiment, the enzyme / substrate reaction emits detectable light proportional to the amount of the target analyte in the sample. Optionally, the enzyme / substrate combination is a luciferase / luciferin. In some embodiments, the emitted light is measured. Optionally, the emitted light is measured using a luminometer. The measured values ​​can then be compared to a standard curve to quantify the amount of the analyte in the biological sample.

[0088] The apparatus and methods described herein can be used to detect the presence or absence of one or more components in a sample. The components may optionally be biological components, including but not limited to proteins (e.g., antibodies, blood proteins, intracellular proteins, membrane proteins, etc.), RNA, DNA, lipids, or others. Optionally, the material to be detected is an organism or a part thereof, optionally bacteria, viruses, fungi, or other organisms. In some embodiments, the organism is bacteria.

[0089] Several aspects of the present invention are illustrated by the following non-limiting examples. These examples are for illustrative purposes only and do not constitute a limitation on any implementation of the invention. However, it should be understood that various changes and modifications can be made without departing from the spirit and scope of the invention.

[0090] Example

[0091] The following examples are provided by way of illustration and are presented in a manner that should be understood by those skilled in the art. These examples are not intended to limit the overall content of this disclosure or the appended claims.

[0092] Example 1:

[0093] In the first set of experiments, the use of the sample storage device for RNA isolation and detection after sample storage was demonstrated. The test sample device was constructed by clamping the sample between two sample membranes, each with a deliquescent layer. A sample containing 5 × 10⁻⁶... 6 A sample of Jurkat cells (a T lymphocyte line) was incubated with 200 μL of vitrification medium containing 600 mM trehalose, 5% glycerol, and 1% Triton X-100 (a nonionic surfactant). The solution was then added to the apparatus. The apparatus containing the sample was stored overnight at 24 °C.

[0094] Then, during RNA isolation using the centrifugation column method, the stored samples underwent thermogravimetric analysis and were compared with freshly cultured cells. The results of the five samples were averaged, and the percentage of mass loss was calculated. The results are shown in Table 1.

[0095] average value Standard error % quality loss 3.69 0.39

[0096] Table 1

[0097] The comparison results are in Figures 5A to 5E As shown in the figure. After RNA isolation, the samples were treated with DNase (deoxyribonuclease), and the RNA was quantified using a Bio Tek Take3 microplate (AGILENT, Santa Clara, California, USA), analyzed by absorbance ratio at 260 nm / 280 nm. The results are as follows. Figure 5A As shown, the RNA concentration isolated from cultured cells was 248.83 ng / μL, while the RNA concentration isolated from cells stored using CaCl2 as a deliquescent agent was 129.49 ng / μL. The RNA recovery rate of CaCl2 vitrified cells was 51% (±4.4%) of the RNA isolated from cultured cells.

[0098] RNA quality was assessed using a fluorescence assay with the QUIBIT RNA kit purchased from Thermo Fisher Scientific (Waltham, MA) to analyze sample degradation. The percentage of small RNA molecules was [not specified]. Figure 5C As shown, the percentage of large RNA molecules is... Figure 5D As shown in the image. Figure 5BThe respective RNA integrity values ​​are shown in the figure. As shown, the RNA integrity value isolated from cultured cells is 9.4, while the RNA integrity value isolated from CaCl2 vitrified cells is 9.53. The results indicate that the quality of RNA vitrified and stored according to the method described herein is almost identical, as is that of freshly obtained RNA. The high-quality RNA isolated from vitrified cells demonstrates that the deliquescence system described herein can yield samples with excellent storage stability.

[0099] RNA quality was confirmed by gel electrophoresis (agarose gel electrophoresis). 0.5 μg of purified RNA, either freshly obtained or vitrified using the apparatus provided herein, was added to each well. Results are in... Figure 5E As shown in the figure. Compared with the control group, this device provides almost the same RNA quality in terms of RNA integrity values, percentage of small RNA, and percentage of large RNA.

[0100] Example 2

[0101] In the second set of experiments, the vitrification of the vitrification medium on the sample film was achieved without direct contact between the sample film and the deliquescent material.

[0102] A specific volume of vitrification medium was loaded onto a 24 mm Ahlstrom sample membrane and stored in a container. The container was placed in a polyester film bag (Mylar) containing CaCl2 desiccant. Four sample volumes (17 μl, 70 μl, 120 μl, and 175 μl) were tested at three time points (days 1, 2, and 3). The weight of the desiccant was recorded before and after storage. Each time point was tested three times.

[0103] Residual moisture was calculated using thermogravimetric analysis. The sum of total volatiles at sample loading and moisture absorbed during measurement equals the sum of the weight gain of the desiccant from the support, the residual moisture on the support, and the weight gain of the desiccant from the storage environment. These results are shown in Table 2.

[0104]

[0105] Table 2

[0106] As shown in the figure, the residual moisture values ​​are all relatively low, reaching their maximum at approximately 24 hours. However, these residual moisture values ​​are based on several assumptions. First, it is assumed that the solution loaded onto the sample membrane completely covers the calculated surface area of ​​the membrane. Second, it is assumed that the punched sample used for thermogravimetric analysis is taken only from the membrane area with the solution loaded. However, this is not strictly true in reality, as the sample may include membrane areas without the solution loaded. This difference leads to lower measured residual moisture values, especially under low sample volume conditions, because areas without the solution loaded do not contribute to the moisture content, thus shifting the results downwards.

[0107] Thermogravimetric analysis was performed at multiple time points (4 hours, 8 hours, 12 hours, 16 hours, and 20 hours) using a loading of 175 μL to determine sufficient drying time with desiccant. These results are shown in Table 3, indicating that 12 hours or longer is sufficient for vitrification.

[0108]

[0109] Table 3

[0110] Vitrification experiments were repeated for 24 hours on 24 mM Ahlstrom membranes using CaCl2 desiccant, with 175 μL of vitrification medium (PBS containing 9.2% BSA, 600 mM trehalose, and 5% glycerol) and 84 μg of luciferase. As a control group, 175 μL of vitrification medium (PBS containing 9.2% BSA, 600 mM trehalose, and 5% glycerol) and 84 μg of luciferase were added to 24 mM Ahlstrom membranes without vitrification.

[0111] The samples were subjected to a stress test at 55°C for one week to simulate accelerated aging conditions. Sample measurements were taken before and after the test. The luciferase control sample showed an increase of 6 mg, while the vitrified sample showed an increase of 150 mg. Results showed a recovery rate of 108.0% for the control sample and 108.3% for the vitrified sample, indicating that the deliquescence system described herein can provide samples with excellent storage stability, similar to that of fresh samples.

[0112] List of items:

[0113] In this first entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device comprising: an outer shell defining a sample chamber configured to contain a biological sample; a deliquescent layer comprising one or more deliquescent materials; and a sample membrane; wherein the deliquescent material is associated with the sample membrane to optionally absorb moisture therefrom by direct contact and / or by fluid communication.

[0114] In the second entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, wherein the deliquescent material is selected from the group consisting of: calcium chloride, magnesium chloride, sodium hydroxide, zinc chloride, ferric chloride, potassium carbonate, potassium phosphate, ferric ammonium citrate, ammonium nitrate, potassium hydroxide, copper sulfate, ammonium persulfate, potassium carbonate, and combinations thereof.

[0115] In the third entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device wherein the deliquescent material is calcium chloride.

[0116] In the fourth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device wherein the sample membrane is a woven material, a spun material, a foam material, a plurality of capillary channels, or a material having a structure capable of promoting fluid capillary action.

[0117] In the fifth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, the sample membrane comprising one or more components of a vitrification medium, optionally wherein said one or more components are vitrified within the sample membrane.

[0118] In the sixth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device wherein the sample membrane comprises trehalose, glycerol, or a combination thereof.

[0119] In the seventh entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, the sample storage device further comprising a polymer layer comprising a vapor-permeable material.

[0120] In the eighth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, wherein the outer casing further includes a base and a cover.

[0121] In the ninth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, wherein the outer casing further includes a hinge connecting the base and the cover.

[0122] In the tenth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device, the sample storage device further comprising a sealing mechanism configured to seal the base and the cover.

[0123] In the eleventh entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device wherein the sealing mechanism is an adhesive tape.

[0124] In the twelfth entry, alone or in combination with any other entry herein, the present invention relates to a sample storage device that also includes a cap.

[0125] In entry thirteen, alone or in combination with any other entry herein, the present invention relates to a sample storage device that also includes a sample collection surface.

[0126] In entry fourteen, alone or in combination with any other entry herein, the present invention relates to a sample storage device wherein a sample collection surface is coupled to a cap.

[0127] In entry 15, alone or in combination with any other entry herein, the present invention relates to a sampling kit comprising: a sample storage device according to any one of the preceding entries; and a sample collection device.

[0128] In the sixteenth entry, alone or in combination with any other entry herein, the present invention relates to a sampling kit, the sampling kit further comprising a bottle containing a vitrified medium.

[0129] In entry seventeen, alone or in combination with any other entry herein, the present invention relates to a method of storing a sample, the method comprising: inserting a sample into or onto a sample membrane of a sample storage device according to any one of claims 1 to 16; and optionally sealing the sample storage device.

[0130] In entry eighteen, alone or in combination with any other entry herein, the present invention relates to a method of storing a sample in which a deliquescent material absorbs moisture from the sample.

[0131] In entry nineteen, alone or in combination with any other entry herein, the present invention relates to a method for storing a sample, wherein the deliquescent material vitrifies the biological sample to produce a storage-stable sample.

[0132] In the twentieth entry, alone or in combination with any other entry herein, the present invention relates to a method for storing a sample, wherein the sample is dried to a moisture residual ratio (MRR) of 0.01 to 0.4.

[0133] In entry 21, alone or in combination with any other entry herein, the present invention relates to a method for storing samples, wherein the MRR is 0.01 to 0.2, or 0.09 to 0.15.

[0134] In entry twenty-two, alone or in combination with any other entry herein, the present invention relates to a method for storing samples, wherein the samples are collected at a location remote from a testing facility.

[0135] In entry twenty-three, alone or in combination with any other entry herein, the present invention relates to a method of storing a sample, the method of storing a sample further comprising transporting the sample storage device to the testing facility.

[0136] In entry twenty-four, alone or in combination with any other entry herein, the present invention relates to a method of storing a sample, the method further comprising restoring the sample to form a restored sample.

[0137] In entry 25, alone or in combination with any other entry herein, the present invention relates to a method for storing a sample, the method further comprising testing the reconstituted sample to detect the presence of an analyte in the reconstituted sample.

[0138] In entry twenty-six, alone or in combination with any other entry herein, the present invention relates to a method for storing a sample, the method further comprising quantitatively detecting the presence of an analyte in the sample.

[0139] In entry twenty-seven, alone or in combination with any other entry herein, the present invention relates to a method for storing a sample, wherein the analyte is a bioanalyte.

[0140] In entry 28, alone or in combination with any other entry herein, the present invention relates to a method for storing samples, wherein the bioanalyte is a cell, bacteria, virus, yeast, protein, antibody, nucleic acid, hormone, fatty acid, steroid, or a combination thereof.

[0141] It should be noted that the terms "substantially" and "about" are used herein to indicate the degree of uncertainty inherent in any quantitative comparison, numerical value, measurement, or other representation. These terms are also used herein to indicate the permissible range of variation of a quantitative representation relative to a given reference value, where such variation does not alter the essential function of the subject matter. The term "substantially" is also used herein to indicate the permissible deviation of a quantitative representation relative to a given reference value, where such deviation does not alter the essential function of the subject matter. Therefore, this term is used to indicate the degree of uncertainty inherent in any quantitative comparison, numerical value, measurement, or other representation, whose arrangement or characteristics, theoretically expected to exhibit a perfectly consistent or perfectly corresponding relationship, may in practice exhibit an imperfectly precise state.

[0142] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for describing particular embodiments only and is not intended to limit the scope of the invention. As used in the specification and appended claims, the singular forms “a,” “an,” and “the” are also intended to include the plural forms, unless the context clearly indicates otherwise.

[0143] In this document, the terms "horizontal" and "vertical" are relative terms only, indicating only a general relative directional relationship and not necessarily indicating perpendicularity. These terms may also be used conveniently to refer to orientations used in the figures, which are used only as a convention and are not intended to be structural features of the illustrated apparatus. The invention and the embodiments described herein can be used with any desired orientation. Furthermore, horizontal and vertical walls generally only need to be intersecting walls, not necessarily perpendicular to each other.

[0144] Note that one or more of the following claims use the term "wherein" as a transitional phrase. To define this art, note that this term is introduced in the claims as an open-ended transitional phrase used to introduce a description of a series of features of the structure, and should be interpreted in a similar manner to the more commonly used open-ended leading term "comprising."

[0145] It should be understood that when a first component is described as "comprising" or "including" a second component, it is conceivable that in some embodiments, the first component may also be "consists of" or "consists essentially of". Furthermore, "consists essentially of" is used in this disclosure to refer to a quantitative value that does not substantially affect the basic and novel features of this disclosure.

[0146] It should be understood that any two quantitative values ​​assigned to a property or measurement parameter can constitute a range of that property or measurement parameter, and all combinations of ranges formed by all the aforementioned quantitative values ​​of a given property or measurement parameter are included within the scope of this invention.

[0147] In addition to the contents shown and described herein, various modifications to the present invention will be apparent to those skilled in the art, and these modifications are also intended to fall within the scope of protection of the appended claims.

[0148] The patents, publications, and applications mentioned in this specification are used to illustrate the technical skill of a person skilled in the art to which this invention pertains. These patents, publications, and applications are incorporated herein by reference as if each patent, publication, or application were separately and individually expressly cited and incorporated herein.

[0149] Although specific embodiments have been described and illustrated herein, it should be understood that various other changes and modifications may be made without departing from the scope of the claimed subject matter. Furthermore, while various aspects of the claimed subject matter have been described herein, these aspects need not be used in combination. Therefore, the appended claims are intended to cover all such changes and modifications within the scope of the claimed subject matter.

Claims

1. A sample storage device, comprising: A shell defining a sample chamber, the sample chamber being configured to contain biological samples; Deliquescent layer, wherein the deliquescent layer comprises one or more deliquescent materials; as well as Sample membrane; in: The deliquescent material is associated with the sample membrane to enable it to absorb moisture from the sample membrane.

2. The sample storage device according to claim 1, wherein, The deliquescent material is selected from the group consisting of: calcium chloride, magnesium chloride, sodium hydroxide, zinc chloride, ferric chloride, potassium carbonate, potassium phosphate, ferric ammonium citrate, ammonium nitrate, potassium hydroxide, copper sulfate, ammonium persulfate, potassium carbonate, and combinations thereof.

3. The sample storage device according to claim 1, wherein, The deliquescent material is calcium chloride.

4. The sample storage device according to claim 1, wherein, The sample membrane is a woven material, a spun material, a foam material, a material with multiple capillary channels, or a material with a structure that can promote fluid capillary action.

5. The sample storage device according to any one of claims 1 to 4, wherein, The sample film contains one or more components of a vitrification medium, wherein optionally the one or more components are vitrified within the sample film.

6. The sample storage device according to claim 5, wherein, The sample membrane contains bovine serum albumin, trehalose, glycerol, or a combination thereof.

7. The sample storage device according to any one of claims 1 to 4, further comprising a polymer layer comprising a vapor-permeable material.

8. The sample storage device according to any one of claims 1 to 4, wherein, The outer shell also includes a base and a cover.

9. The sample storage device according to claim 8, wherein, The outer casing also includes a hinge connecting the base and the cover.

10. The sample storage device according to claim 8, further comprising a sealing mechanism configured to seal the base and the cover.

11. The sample storage device according to claim 10, wherein, The sealing mechanism is a tape.

12. The sample storage device according to any one of claims 1 to 4, further comprising a cap.

13. The sample storage device according to claim 12 further includes a sample collection surface.

14. The sample storage device according to claim 13, wherein, The sample collection surface is connected to the cap.

15. The sample storage device according to any one of claims 1 to 4, wherein, The deliquescent material is associated with the sample membrane through direct contact and / or through fluid communication.

16. A sampling kit comprising: The sample storage device according to any one of claims 1 to 4; as well as Sample collection device.

17. The sampling kit of claim 16 further includes a bottle containing a vitrified medium.

18. A method for storing a sample, comprising: Insert the sample into or onto the sample membrane of the sample storage device according to any one of claims 1 to 4; as well as The sample storage device may be optionally sealed.

19. The method for storing a sample according to claim 18, wherein, The deliquescent material absorbs moisture from the sample.

20. The method for storing a sample according to claim 18, wherein, The deliquescent material vitrifies the biological sample to produce a storage-stable sample.

21. The method for storing samples according to claim 18, wherein, The samples were dried to a residual moisture ratio (MRR) of 0.01 to 0.

4.

22. The method for storing samples according to claim 21, wherein, The MRR is 0.01 to 0.2, or 0.09 to 0.

15.

23. The method according to claim 18, wherein, The samples were collected at a location far from the testing facility.

24. The method of claim 23, further comprising transporting the sample storage device to the testing facility.

25. The method of claim 24, further comprising restoring the sample to form a restored sample.

26. The method of claim 25 further includes detecting the restored sample to detect the presence of an analyte in the sample.

27. The method of claim 26 further includes quantitatively detecting the presence of the analyte in the sample.

28. The method according to claim 26, wherein, The analyte is a bioanalyte.

29. The method according to claim 28, wherein, The bioanalytes are cells, bacteria, viruses, yeast, proteins, antibodies, nucleic acids, hormones, fatty acids, steroids, or combinations thereof.

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