Nanoscale Imaging of RNA with Expansion Microscopy
Wassie, Asmamaw T.
Cote, Allison J.
Boyden, Edward S.Note: Order does not necessarily reflect citation order of authors.
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CitationChen, F., A. T. Wassie, A. J. Cote, A. Sinha, S. Alon, S. Asano, E. R. Daugharthy, et al. 2016. “Nanoscale Imaging of RNA with Expansion Microscopy.” Nature methods 13 (8): 679-684. doi:10.1038/nmeth.3899. http://dx.doi.org/10.1038/nmeth.3899.
AbstractThe 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 (ExM) 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, post-expansion, fluorescent in situ hybridization (FISH) imaging of RNA can be performed with high yield and specificity, with single molecule precision, in both cultured cells and intact brain tissue. Expansion FISH (ExFISH) de-crowds RNAs and supports amplification of single molecule signals (i.e., via hybridization chain reaction (HCR)) 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.
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