Nanoscale imaging of proteins and nucleic acids via expansion microscopy
AssigneeMassachusetts Institute of Technology
InventorsAsmamaw Wassie, Fei Chen, Edward Stuart Boyden, Shahar Alon, George Church, Evan Daugharthy
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Abstract
The invention enables in situ genomic and transcriptomic assessment of nucleic acids to be conducted in biological specimens that have been physically expanded. The invention leverages the techniques for expansion microscopy (ExM) to provide new methods for in situ genomic and transcriptomic assessment of nucleic in a new process referred to herein as “expansion fluorescent in situ hybridization” (ExFISH).
Claims13
A method for in situ genomic and transcriptomic assessment of target nucleic acids present in a biological sample comprising the steps of:
- a) treating the biological sample with a small molecule linker comprising a chemical reactive group capable of covalently binding at least one target nucleic acid and a chemical group that can be incorporated into the a swellable material;
- b) permeating the biological sample with a composition comprising precursors of a swellable material;
- c) initiating polymerization of the precursors to form a swellable material, wherein the small molecule linker is bound to the at least one target nucleic acid in the biological sample and to the swellable material;
- d). subjecting the biological sample to a physical disruption method;
- e). swelling the swellable material to form an expanded biological sample;
- f). providing at least one oligonucleotide complementary to the at least one target nucleic acid, wherein the at least one oligonucleotide hybridizes to the at least one target nucleic acid;
- and
- g). genomically or transcriptomically assessing the expanded biological sample.
Dependencies are inferred from the claim text.