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 carrier substrate, thereby immobilizing the tissue section onto said carrier substrate and generating a sample-carrier construct, wherein said carrier substrate comprises a first adhesion strength;
b) contacting the tissue section of the sample-carrier construct with a receiving substrate to generate an immobilized tissue section, wherein said receiving substrate comprises a second adhesion strength, wherein the second adhesion strength is greater than the first adhesion strength;
c) removing the carrier substrate from the immobilized tissue section;
d) permeabilizing the immobilized tissue section; and
e) contacting said biomolecule in said tissue section with a detection agent, thereby detecting the biomolecule in the tissue section, wherein the detection agent comprises a fluorophore.
2 . The method of claim 1 , wherein step c) occurs prior to step e).
3 . The method of claim 1 , wherein step c) occurs prior to step d).
4 . The method of claim 1 , wherein generating a sample-carrier construct comprises forming a plurality of non-covalent bonds between the tissue section and the carrier substrate.
5 . The method of claim 1 , wherein the carrier substrate comprises water molecules attached to the surface of said carrier substrate.
6 . The method of claim 1 , wherein the carrier substrate comprises a compression modulus greater than about 100 kPa.
7 . The method of claim 1 , wherein generating an immobilized tissue section comprises forming a plurality of covalent bonds between the tissue section and the receiving substrate.
8 . The method of claim 1 , wherein the receiving 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 sequence, carbohydrate, or protein.
10 . The method of claim 1 , wherein the biomolecule is a nucleic acid sequence.
11 . The method of claim 10 , further comprising amplifying the nucleic acid sequence to generate amplification products.
12 . The method of claim 11 , further comprising detecting the amplification products.
13 . The method of claim 1 , wherein the 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 . The method of claim 10 , wherein step e) comprises hybridizing a sequencing primer to the biomolecule and sequencing the biomolecule.
18 .- 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 carrier 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 carrier substrate comprises agarose, amylose, or amylopectin.
28 . (canceled)
29 . The method of claim 1 , wherein the carrier substrate further comprises a support scaffold, wherein said support scaffold is a thermoplastic elastomer.
30 . The method of claim 1 , wherein the carrier substrate comprises a Young's modulus of about 5 kPa to about 30 kPa.
31 . (canceled)
32 . The method of claim 1 , wherein the carrier substrate comprises about 80% to about 99% water.
33 . The method of claim 1 , wherein the receiving substrate comprises a functionalized glass surface or a functionalized plastic surface.
34 . The method of claim 1 , wherein prior to contacting the tissue section with the receiving 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 about 25° C.
38 .- 39 . (canceled)
40 . The method of claim 1 , wherein removing the carrier substrate comprises physically removing, thermally removing, chemically removing, or enzymatically removing.
41 .- 98 . (canceled)
99 . The method of claim 1 , wherein the sample-carrier construct comprises substantially uniform adhesion between the tissue section and the carrier substrate.
100 . The method of claim 1 , wherein the carrier substrate comprises a polymerized hydrogel.
101 . The method of claim 1 , wherein substantially all of the tissue section is immobilized to the receiving substrate.
102 . The method of claim 1 , wherein prior to step a), the carrier substrate is solid or semi-solid.
103 . The method of claim 40 , wherein physically removing said carrier substrate comprises mechanically pulling or lifting thereby detaching said carrier substrate.