Nanoscale imaging of RNA with expansion microscopy

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

Research output: Contribution to journalArticlepeer-review

244 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)679-684
Number of pages6
JournalNature Methods
Volume13
Issue number8
DOIs
StatePublished - Aug 2016
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2016 Nature America, Inc.

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