Title |
Expansion Microscopy: Protocols for Imaging Proteins and RNA in Cells and Tissues
|
---|---|
Published in |
Current protocols in cell biology, August 2018
|
DOI | 10.1002/cpcb.56 |
Pubmed ID | |
Authors |
Shoh M. Asano, Ruixuan Gao, Asmamaw T. Wassie, Paul W. Tillberg, Fei Chen, Edward S. Boyden |
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. © 2018 by John Wiley & Sons, Inc. |
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