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URL: https://dx.doi.org/DOI:10.17504/protocols.io.n6adhae

Authors: Fei Chen, Asmamaw T Wassie, Allison J Cote, Anubhav Sinha, Shahar Alon, Shoh Asano, Evan R Daugharthy, Jae-Byum Chang, Adam Marblestone, George M Church, Arjun Raj, Edward S Boyden

Group: Human Cell Atlas Method Development Community

Summary: The ability to image RNA identity and location with nanoscale precision in intact tissues is of great interest for defining cell types and states in normal and pathological biological settings. Here, we present a strategy for expansion microscopy of RNA. We developed a small-molecule linker that enables RNA to be covalently attached to a swellable polyelectrolyte gel synthesized throughout a biological specimen. Then, postexpansion, fluorescent in situ hybridization (FISH) imaging of RNA can be performed with high yield and specificity as well as single-molecule precision in both cultured cells and intact brain tissue. Expansion FISH (ExFISH) separates RNAs and supports amplification of single-molecule signals (i.e., via hybridization chain reaction) as well as multiplexed RNA FISH readout. ExFISH thus enables super-resolution imaging of RNA structure and location with diffraction-limited microscopes in thick specimens, such as intact brain tissue and other tissues of importance to biology and medicine.

Affiliations: Massachusetts Institute of Technology, Massachusetts Institute of Technology, University of Pennsylvania, Massachusetts Institute of Technology, Massachusetts Institute of Technology, Massachusetts Institute of Technology, Harvard University, Massachusetts Institute of Technology, Massachusetts Institute of Technology, Harvard University, University of Pennsylvania, Massachusetts Institute of Technology

External URL: http://expansionmicroscopy.org/ExFISHV1.0.docx

Version: 1

Publication Date: 2018

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