ExFISH - Cultured Cells v1

• DOI: 10.17504/protocols.io.n32dgqe
• Publication Date: 2018-04-24T18:42:31Z
• 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
• Keywords: Linker

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