Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues
Name
nihms960054.pdf
Description
Accepted version
Size
2.17 MB
Format
Adobe PDF
Checksum (MD5)
abf0d3bce6b7f9f0afe212576cf41c6b
Author(s)
Asano, Shoh M.
Gao, Ruixuan
Wassie, Asmamaw T.
Tillberg, Paul W.
Chen, Fei
Boyden, Edward
Date Issued
August 2018
Journal
Current Protocols in Cell Biology
Publisher
Wiley
Citation
Asano, Shoh M. et al. "Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues." Current Protocols in Cell Biology 80, 1 (September 2018): e56 © 2018 John Wiley & Sons, Inc.
Version
Author's final manuscript
Abstract
Expansion microscopy (ExM) is a recently developed technique that enables nanoscale-resolution imaging of preserved cells and tissues on conventional diffraction-limited microscopes via isotropic physical expansion of the specimens before imaging. In ExM, biomolecules and/or fluorescent labels in the specimen are linked to a dense, expandable polymer matrix synthesized evenly throughout the specimen, which undergoes 3-dimensional expansion by ∼4.5 fold linearly when immersed in water. Since our first report, versions of ExM optimized for visualization of proteins, RNA, and other biomolecules have emerged. Here we describe best-practice, step-by-step ExM protocols for performing analysis of proteins (protein retention ExM, or proExM) as well as RNAs (expansion fluorescence in situ hybridization, or ExFISH), using chemicals and hardware found in a typical biology lab. Furthermore, a detailed protocol for handling and mounting expanded samples and for imaging them with confocal and light-sheet microscopes is provided.
MIT Department
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Media Laboratory
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Terms of Use
Creative Commons Attribution-Noncommercial-Share Alike
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1002/cpcb.56
