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 . A method of detecting a biomolecule in a tissue section, said method comprising:
a) contacting the tissue section with a first solid substrate, thereby immobilizing the tissue section onto the first solid substrate and generating a sample-carrier construct, wherein the tissue section is bound to the first solid substrate at a first adhesion strength;
b) contacting the tissue section of the sample-carrier construct with a second solid substrate to generate an immobilized tissue section, wherein the tissue section is bound to the second solid substrate at a second adhesion strength, wherein the second adhesion strength is greater than the first adhesion strength;
c) removing the first solid substrate from the immobilized tissue section;
d) contacting a biomolecule in said tissue section with a labeled detection agent; and
e) detecting a light emission from the labeled detection agent, thereby detecting the biomolecule in the tissue section.
2 .- 4 . (canceled)
5 . The method of claim 1 , wherein the first solid substrate comprises water molecules attached to the surface of said first solid substrate.
6 . The method of claim 1 , wherein the first solid substrate comprises a compression modulus greater than about 100 kPa.
7 . (canceled)
8 . The method of claim 1 , wherein the second solid substrate 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.
9 . The method of claim 1 , wherein the biomolecule is a nucleic acid molecule, carbohydrate, or protein.
10 . The method of claim 1 , wherein the biomolecule is a nucleic acid molecule.
11 . The method of claim 10 , further comprising amplifying the nucleic acid molecule to generate amplification products.
12 . The method of claim 11 , further comprising detecting the amplification products.
13 . The method of claim 1 , wherein the labeled detection agent comprises a protein-specific binding agent.
14 .- 15 . (canceled)
16 . The method of claim 1 , further comprising digesting the tissue section by contacting the sample-carrier construct with an endopeptidase.
17 .- 24 . (canceled)
25 . The method of claim 1 , wherein the thickness of the tissue section is about 1 μm to about 20 μm.
26 . The method of claim 1 , wherein the first solid substrate comprises agarose, amylose, amylopectin, alginate, gelatin, cellulose, polyolefin, polyethylene glycol, polyvinyl alcohol, and/or acrylate polymers and copolymers thereof.
27 . The method of claim 1 , wherein the first solid substrate comprises agarose, amylose, or amylopectin.
28 . (canceled)
29 . The method of claim 1 , wherein the first solid substrate further comprises a support scaffold, wherein said support scaffold is a thermoplastic elastomer.
30 . The method of claim 1 , wherein the first solid substrate comprises a Young's modulus of about 5 kPa to about 30 kPa.
31 . The method of claim 1 , wherein the sample-carrier construct comprises interfacial water, wherein the interfacial water is between the first solid substrate and the tissue section.
32 .- 33 . (canceled)
34 . The method of claim 1 , wherein prior to contacting the tissue section with the second solid substrate, the sample-carrier construct is stored for one or more days.
35 .- 36 . (canceled)
37 . The method of claim 34 , wherein the sample-carrier construct is stored at less than 25° C.
38 .- 39 . (canceled)
40 . The method of claim 1 , wherein removing the first solid substrate comprises physically removing, thermally removing, chemically removing, or enzymatically removing.
41 . A method of detecting a nucleic acid in a tissue section, said method comprising:
a) immobilizing the tissue section onto a hydrogel carrier substrate to generate a sample-carrier construct;
b) contacting the tissue section of the sample-carrier construct with a receiving substrate to generate an immobilized tissue section;
c) removing the hydrogel carrier substrate from the immobilized tissue section;
d) hybridizing a polynucleotide probe to the nucleic acid molecule and amplifying the polynucleotide probe to generate an amplification product;
and
e) hybridizing a primer to the amplification product and incorporating a labeled nucleotide and detecting the labeled nucleotide thereby detecting the biomolecule in the tissue section.
42 .- 98 . (canceled)