Devices and methods for tissue and cell staining

Abstract

Devices and systems are provided herein relating to a novel and rapid assay for tissue staining. Methods for using the devices and systems for analyzing tissue samples are also disclosed.

We claim: 
     
         1 . A device for analyzing a tissue sample, comprising:
 a first plate; a second plate; a plurality of spacers; and a staining liquid, wherein:
 (i) the first and second plates are movable relative to each other into different configurations, including an open configuration and a closed configuration; 
 (ii) the first plate is flexible and comprises the plurality spacers on the first plate; 
 (iii) each of the plates has, on its respective inner surface, a sample contact area for contacting the staining liquid and/or a tissue sample containing or suspected of containing a target analyte; 
 (iv) the plurality of spacers have a predetermined substantially uniform height that is 200 microns or less, and a predetermined inter-spacer distance; 
 (v) at least one of the plurality of spacers is inside the sample contact area; and 
 (vi) the staining liquid is deposited on (a) one or both of the plates in the open configuration or (b) the tissue sample; 
 wherein the thickness of the first plate times the Young's modulus of the first plate is in the range of 60 to 750 GPa-μm; 
 wherein the fourth power of the inter-spacer-distance (ISD) of the plurality of spacers divided by the thickness (h) and the Young's modulus (E) of the flexible plate (ISD 4 /(hE)) is 5×10 6  μm 3 /GPa or less; 
 wherein the open configuration is the configuration, in which: the two plates are partially or entirely separated apart, the spacing between the plates is not regulated by the plurality of spacers, and the tissue sample is deposited on one or both of the plates; and 
 wherein the closed configuration is the configuration that is configured after the open configuration; and in the closed configuration: at least part of the tissue sample and a layer of at least part of the staining liquid are between the two plates, wherein the thickness of the layer of at least part of the staining liquid is regulated by the plates, the tissue sample, and the plurality of spacers. 
   
     
     
         2 . The device of  claim 1 , further comprising a stain agent coated on one or both of the sample contact areas of the plates, wherein the staining liquid comprises a transfer solution, and wherein the stain agent is configured, upon contacting the transfer solution, to be dissolved in the transfer solution to form a liquid that stains the tissue sample. 
     
     
         3 . A method for analyzing a tissue sample, comprising:
 (a) obtaining a tissue sample containing or suspected of containing a target analyte;   (b) obtaining the device of  claim 1 ;   (c) depositing, when the plates are in the open configuration, (i) the tissue sample on one or both of the plates and (ii) the staining liquid on the tissue sample or on one or both of the plates;   (d) after (c), bringing the two plates together and pressing the plates into the closed configuration; and   (e) analyzing, without washing the tissue sample, the target analyte in the tissue sample when the plates are in the closed configuration.   
     
     
         4 . A method for analyzing a tissue sample, comprising:
 (a) obtaining a tissue sample containing or suspected of containing a target analyte;   (b) obtaining the device of  claim 2 ;   (c) depositing, when the plates are in the open configuration, (i) the tissue sample on one or both of the plates and (ii) the transfer solution on the tissue sample or on one or both of the plates;   (d) after (c), bringing the two plates together and pressing the plates into the closed configuration; and   (e) analyzing, without washing the tissue sample, the target analyte in the tissue sample when the plates are in the closed configuration.   
     
     
         5 . The method of  claim 3 , wherein the step (e) uses one or more imagers that image the tissue sample generating one or more images. 
     
     
         6 . The method of  claim 4 , wherein the step (e) uses one or more imagers that image the tissue sample generating one or more images. 
     
     
         7 . A system for analyzing a tissue sample, comprising:
 (a) the device of  claim 1 ; and   (b) a detector, wherein the detector detects signals of the target analyte in the tissue sample.   
     
     
         8 . A smartphone system for tissue analysis assay, comprising:
 (a) the device of  claim 1 ; and   (b) a mobile communication device that comprises:
 (i) one or a plurality of cameras for detecting and/or imaging the tissue sample, and 
 (ii) electronics, signal processors, hardware and software for receiving and/or processing the detected signal and/or the image of the tissue sample and for remote communication. 
   
     
     
         9 . The method of  claim 3 , wherein one or both of the plates is configured such that the tissue sample can be dried thereon at the open configuration, and wherein the tissue sample comprises bodily fluid selected from the group consisting of amniotic fluid, aqueous humour, vitreous humour, blood, breast milk, cerebrospinal fluid (CSF), cerumen (earwax), chyle, chime, endolymph, perilymph, feces, breath, gastric acid, gastric juice, lymph, mucus, pericardial fluid, peritoneal fluid, pleural fluid, pus, rheum, saliva, exhaled breath condensates, sebum, semen, sputum, sweat, synovial fluid, tears, vomit, urine, and any combination thereof. 
     
     
         10 . The method of  claim 3 , wherein the tissue sample is mucus. 
     
     
         11 . The device of  claim 1 , wherein the staining liquid comprises a permeabilizing agent capable of permeabilizing cells in the tissue sample that contain the target analyte. 
     
     
         12 . The device of  claim 1 , wherein the staining liquid comprises a detection agent that specifically labels the target analyte in the tissue sample. 
     
     
         13 . The device of  claim 1 , wherein the sample contact area of one or both plates comprise a storage site that contains a detection agent that specifically labels the target analyte in the tissue sample. 
     
     
         14 . The device of  claim 1 , wherein the staining liquid comprises a the detection agent, wherein the detection agent comprises a compound selected from the group consisting of: Acid fuchsin, Alcian blue 8 GX, Alizarin red S, Aniline blue WS, Auramine O, Azocarmine B, Azocarmine G, Azure A, Azure B, Azure C, Basic fuchsine, Bismarck brown Y, Brilliant cresyl blue, Brilliant green, Carmine, Chlorazol black E, Congo red, C.I. Cresyl violet, Crystal violet, Darrow red, Eosin B, Eosin Y, Erythrosin, Ethyl eosin, Ethyl green, Fast green F C F, Fluorescein Isothiocyanate, Giemsa Stain, Hematoxylin, Hematoxylin & Eosin, Indigo carmine, Janus green B, Jenner stain 1899, Light green SF, Malachite green, Martius yellow, Methyl orange, Methyl violet 2B, Methylene blue, Methylene blue, Methylene violet, Neutral red, Nigrosin, Nile blue A, Nuclear fast red, Oil Red, Orange G, Orange II, Orcein, Pararosaniline, Phloxin B, Protargol S, Pyronine B, Pyronine, Resazurin, Rose Bengal, Safranine O, Sudan black B, Sudan III, Sudan IV, Tetrachrome stain, Thionine, Toluidine blue, Weigert, Wright stain, and any combination thereof. 
     
     
         15 . The device of  claim 1 , wherein one or both of the plates further comprise, on the respective sample contact area, a cell viability dye selected from the group consisting of: Propidium Iodide, 7-AAD, Trypan blue, Calcein Violet AM, Calcein AM, Fixable Viability Dyes, nucleic acid dyes, Resazurin and Formazan (MTT/XTT) and other mitochondrial dyes, and any combination thereof. 
     
     
         16 . The device of  claim 1  or  2 , wherein the staining liquid comprises a detection agent, wherein the detection agent comprises an antibody that specifically binds to an antigen on the target analyte in the tissue sample. 
     
     
         17 . The device of  claim 1  or  2 , wherein the staining liquid comprises a detection agent, wherein the detection agent comprises an oligonucleotide probe that specifically binds to DNA and/or RNA in the tissue sample. 
     
     
         18 . The device of  claim 1  or  2 , wherein the staining liquid comprises a detection agent that is labeled with a reporter molecule, wherein the reporter molecule provides a detectable signal to be read and analyzed. 
     
     
         19 . The method of  claim 3  or  4 , wherein the analyzing detects a detectable signal that is selected from the group consisting of:
 (i) luminescence selected from photoluminescence, electroluminescence, and electrochemiluminescence, 
 (ii) light absorption, reflection, transmission, diffraction, scattering, or diffusion, 
 (iii) surface Raman scattering, 
 (iv) electrical impedance selected from resistance, capacitance, and inductance, 
 (v) magnetic relaxivity and 
 (vi) a combination thereof. 
 
     
     
         20 . The method of  claim 3 , wherein the staining liquid includes a transfer solution. 
     
     
         21 . The device of  claim 1 , wherein at least a portion of the inner surface of one plate or both plates is hydrophilic. 
     
     
         22 . The device of  claim 1 , wherein the inter spacer distance is periodic. 
     
     
         23 . The device of  claim 1 , further comprising a hinge that connects the first plate and the second plate, and the plates can rotate around the hinge, wherein the hinge is made from a metallic material 
     
     
         24 . The device of  claim 1  wherein the inter spacer distance is equal or less than 120 μm. 
     
     
         25 . The device of  claim 1  wherein the spacers are arranged periodically and the inter spacer distance is equal or less than 120 μm. 
     
     
         26 . The device of  claim 1 , wherein the thickness of the first plate times the Young's modulus of the first plate is in the range of 60 to 550 GPa-μm, and wherein the fourth power of the inter-spacer-distance (ISD) divided by the thickness (h) and the Young's modulus (E) of the first plate (ISD 4 /(hE)) is 5×10 5  μm 3 /GPa or less. 
     
     
         27 . The method of  claim 3 , wherein the tissue sample has a thickness of 2 μm to 6 μm. 
     
     
         28 . The method of  claim 3 , wherein the plurality of spacers have a shape of pillar, and the pillar has substantially uniform cross-section. 
     
     
         29 . The method of  claim 3 , wherein the tissue sample is for the detection, purification and quantification of chemical compounds or biomolecules that correlates with the stage of certain diseases. 
     
     
         30 . The method of  claim 3 , wherein the tissue sample is related to infectious disease, parasitic disease, injuries, cardiovascular disease, cancer, mental disorders, neuropsychiatric disorders, pulmonary diseases, renal diseases, or other organic diseases. 
     
     
         31 . The method of  claim 3 , wherein the tissue sample is related to the detection and quantification of specific DNA or RNA from bio-samples. 
     
     
         32 . The method of  claim 3 , wherein the tissue sample is related to the sequencing and comparing of genetic sequences in DNA in the chromosomes or mitochondria for genome analysis. 
     
     
         33 . The method of  claim 3 , wherein the tissue sample comprises cells, bodily fluid, or stool. 
     
     
         34 . The method of  claim 3 , wherein the target analyte is a protein, peptides, nucleic acids, synthetic compounds, or inorganic compounds. 
     
     
         35 . The method of  claim 3 , wherein the tissue sample is a sample in the field of human, veterinary, agriculture, foods, environments, or drug testing. 
     
     
         36 . The method of  claim 3 , wherein the tissue sample is a biological sample selected from the group consisting of blood, serum, plasma, a nasal swab, a nasopharyngeal wash, saliva, urine, gastric fluid, spinal fluid, tears, stool, mucus, sweat, earwax, oil, a glandular secretion, cerebral spinal fluid, tissue, semen, vaginal fluid, interstitial fluids derived from tumorous tissue, ocular fluids, spinal fluid, a throat swab, breath, hair, finger nails, skin, biopsy, placental fluid, amniotic fluid, cord blood, lymphatic fluids, cavity fluids, sputum, pus, microbiota, meconium, breast milk, exhaled condensate nasopharyngeal wash, nasal swab, throat swab, stool samples, hair, finger nail, ear wax, breath, connective tissue, muscle tissue, nervous tissue, epithelial tissue, cartilage, cancerous sample, and bone. 
     
     
         37 . The device of  claim 1 , wherein the staining liquid comprises: a fixative capable of fixing the tissue sample; a blocking agent; a deparaffinizing agent capable of removing paraffin in the tissue sample; a permeabilizing agent capable of permeabilizing cells in the tissue sample that contain the target analyte; an antigen retrieval agent capable of facilitating retrieval of antigen; a detection agent that specifically labels the target analyte in the tissue sample; a compound; a detection agent comprising an antibody that specifically binds to an antigen on the target analyte in the tissue sample; a detection agent comprising an oligonucleotide probe that specifically binds to DNA and/or RNA in the tissue sample; a detection agent that is labeled with a reporter molecule that provides a detectable signal to be read and analyzed; or any combination thereof. 
     
     
         38 . The device of  claim 2 , wherein the stain agent comprises: a detection agent that specifically labels the target analyte in the tissue sample; a compound; a detection agent comprising an antibody that specifically binds to an antigen on the target analyte in the tissue sample; a detection agent comprising an oligonucleotide probe that specifically binds to DNA and/or RNA in the tissue sample; a detection agent that is labeled with a reporter molecule that provides a detectable signal to be read and analyzed; or any combination thereof. 
     
     
         39 . The device of  claim 1 , wherein the sample contact area of one or both plates comprises a storage site that contains a blocking agent; a deparaffinizing agent capable of removing paraffin in the tissue sample; a permeabilizing agent capable of permeabilizing cells in the tissue sample that contain the target analyte; an antigen retrieval agent capable of facilitating retrieval of antigen; a detection agent that specifically labels the target analyte in the tissue sample; or any combination thereof. 
     
     
         40 . The method of  claim 3 , wherein the staining liquid comprises oligonucleotide and/or RNA probes for in situ hybridization stain. 
     
     
         41 . The method of  claim 3 , wherein the analyzing comprises a use of light. 
     
     
         42 . The device of  claim 1 , wherein the plurality of spacers has a height of 10 μm. 
     
     
         43 . The device of  claim 1 , wherein the plurality of spacers has a height selected from 2 μm to 6 μm. 
     
     
         44 . The device of  claim 1 , wherein the staining liquid comprising an immunohistochemical (IHC) stain agent. 
     
     
         45 . The device of  claim 1 , wherein the staining liquid comprising a nucleic acid stain agent. 
     
     
         46 . The device of  claim 1 , wherein the staining liquid comprises an H&E stain agent. 
     
     
         47 . The device of  claim 1 , wherein the staining liquid comprises a stain agent that modifies the color of a specimen. 
     
     
         48 . The device of  claim 1 , wherein the staining liquid comprises an immunohistochemical (IHC) stain agent; a nucleic acid stain agent; an H&E stain agent; a stain agent that modifies the color of a specimen; or any combination thereof. 
     
     
         49 . The method of  claim 3 , wherein the analyzing comprises a machine learning. 
     
     
         50 . The device of  claim 1 , wherein the plurality of spacers function as a location marker, a scale marker, an imaging marker, or any combination of thereof. 
     
     
         51 . The device of  claim 1 , wherein one or both plates comprise a location marker, a scale marker, an imaging marker, or any combination of thereof. 
     
     
         52 . The method of  claim 3 , wherein the analyzing is for checking cell viability. 
     
     
         53 . The method of  claim 3 , wherein the sample is mucus, sputum, nasal swab, or throat swab. 
     
     
         54 . The method of  claim 4 , wherein the sample is mucus, sputum, nasal swab, or throat swab. 
     
     
         55 . The method of  claim 3 , wherein the targeted analyte is nucleic acid, and wherein the staining solution comprises a nucleic acid detection agent connected to a reporter that gives a fluorescent signal. 
     
     
         56 . The method of  claim 4 , wherein the targeted analyte is nucleic acid, and wherein the staining solution comprises a nucleic acid detection agent connected to a reporter that gives a fluorescent signal. 
     
     
         57 . The device of  claim 1 , wherein the staining liquid comprises a detection agent that has a reporter comprising fluorescent molecules. 
     
     
         58 . The device of  claim 2 , wherein the staining liquid comprises a detection agent that has a reporter comprising fluorescent molecules.