MANIPULATING AND DETECTING BIOLOGICAL SAMPLES
Disclosed herein, inter alia, are compositions and methods for efficient transfer and analyses of cellular material, tissue samples, such as tissue sections, using carrier substrates.
1 - 64 . (canceled)
65 . A method of obtaining an image of a portion of a tissue section, said method comprising:
A) immobilizing the tissue section onto a hydrogel carrier substrate to generate a sample-carrier construct comprising the carrier substrate and the tissue section;
B) removing a portion of the sample-carrier construct, wherein the portion comprises a portion of the carrier substrate and a portion of the tissue section;
C) contacting the tissue section of the portion of the sample-carrier construct with a receiving substrate to generate an immobilized tissue section;
D) removing the hydrogel carrier substrate from the immobilized tissue section; and
E) imaging the tissue section, thereby obtaining an image of a portion of the tissue section.
66 . The method of claim 65 , wherein prior to step E), the method further comprises permeabilizing the immobilized tissue section.
67 . The method of claim 65 , wherein prior to step E), the method does not comprise permeabilizing the immobilized tissue section.
68 . The method of claim 65 , wherein step E) comprises phase-contrast microscopy, bright-field microscopy, Nomarski differential-interference-contrast microscopy, dark field microscopy, electron microscopy, or cryo-electron microscopy.
69 . The method of claim 65 , wherein prior to step E), the method further comprises contacting the immobilized tissue section with one or more imaging reagents or stains.
70 . The method of claim 69 , wherein the one or more imaging reagents or stains comprise hematoxylin and eosin (H&E) staining reagents.
71 . The method of claim 69 , wherein the one or more imaging reagents or stains comprise phase-contrast microscopy, bright-field microscopy, Nomarski differential-interference-contrast microscopy, or dark field microscopy imaging reagents.
72 . The method of claim 69 , wherein the one or more imaging reagents or stains comprise electron microscopy or cryo-electron microscopy imaging reagents.
73 . The method of claim 65 , wherein the thickness of the tissue section is about 1 µm to about 20 µm.
74 - 75 . (canceled)
76 . The method of 65 , wherein the tissue section is embedded in an embedding material comprising paraffin wax, polyepoxide polymer, polyacrylic polymer, agar, gelatin, celloidin, cryogel, optimal cutting temperature (OCT) compositions, glycols, or a combination thereof.
77 . The method of claim 76 , further comprising removing the embedding material.
78 . The method of claim 76 , further comprising removing the embedding material prior to step C).
79 . The method of claim 65 , wherein the hydrogel carrier substrate comprises agarose, amylose, amylopectin, alginate, gelatin, cellulose, polyolefin, polyethylene glycol, polyvinyl alcohol, and/or acrylate polymers and copolymers thereof.
80 . The method of claim 65 , wherein the hydrogel carrier substrate comprises agarose, amylose, or amylopectin.
81 . The method of claim 65 , wherein the hydrogel carrier substrate comprises less than about 5% agarose.
82 . The method of claim 65 , wherein the hydrogel carrier substrate further comprises a support scaffold.
83 . The method of claim 82 , wherein the support scaffold comprises a thermoplastic elastomer.
84 . (canceled)
85 . The method of claim 65 , wherein the hydrogel carrier substrate comprises a Young’s modulus of about 5 kPa to about 30 kPa.
86 . The method of claim 65 , wherein the sample-carrier construct comprises interfacial water, wherein the interfacial water is between the carrier substrate and the tissue section.
87 . The method of claim 65 , wherein the hydrogel carrier substrate comprises about 80% to about 99% water.
88 . The method of claim 65 , wherein the receiving substrate comprises a functionalized glass surface or a functionalized plastic surface.
89 . The method of claim 88 , wherein the functionalized glass surface comprises (3-aminopropyl)triethoxysilane (APTES), (3-Aminopropyl)trimethoxysilane (APTMS), γ-Aminopropylsilatrane (APS), N-(6-aminohexyl)aminomethyltriethoxysilane (AHAMTES), polyethylenimine (PEI), 5,6-epoxyhexyltriethoxysilane, or triethoxysilylbutyraldehyde, or a combination thereof.
90 . The method of claim 65 , wherein prior to step C), the sample-carrier construct is stored for one or more days.
91 - 92 . (canceled)
93 . The method of claim 90 , wherein the sample-carrier construct is stored at less than about 25° C.
94 - 95 . (canceled)
96 . The method of claim 65 , wherein step D) comprises physically removing, thermally removing, chemically removing, or enzymatically removing.
97 - 98 . (canceled)
99 . The method of claim 65 , wherein the sample-carrier construct comprises substantially uniform adhesion between the tissue section and the carrier substrate.
100 . The method of claim 65 , wherein the tissue section is immobilized onto the surface of the hydrogel carrier substrate.
101 . The method of claim 65 , wherein substantially all of the tissue section of the portion is immobilized to the receiving substrate.
102 . The method of claim 65 , wherein prior to step A), the hydrogel carrier substrate is solid or semi-solid.
103 . A method of immobilizing a portion of a tissue section to a receiving substrate, wherein the tissue section comprises a thickness of about 1 µm to about 50 µm, said method comprising: contacting the tissue section with a hydrogel carrier substrate to generate a sample-carrier construct comprising the carrier substrate and the tissue section; removing a portion of the sample-carrier construct, wherein the portion comprises a portion of the carrier substrate and a portion of the tissue section; contacting the tissue section of the portion of the sample-carrier construct with the receiving substrate thereby immobilizing the tissue section to the receiving substrate.