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The Basics of Expansion Microscopy

Authors
  • Klimas, Aleksandra
  • Gallagher, Brendan
  • Zhao, Yongxin
Type
Published Article
Journal
Current protocols in cytometry
Publication Date
Dec 01, 2019
Volume
91
Issue
1
Identifiers
DOI: 10.1002/cpcy.67
PMID: 31763769
PMCID: PMC6880804
Source
PubMed Central
Keywords
Disciplines
  • Article
License
Unknown

Abstract

Optical imaging techniques are often used in neuroscience to understand brain function and discern disease pathogenesis. However, the optical diffraction limit precludes conventional optical imaging approaches from resolving nanoscopic structures with feature sizes smaller than 300 nm. Expansion microscopy (ExM) circumvents this limit by physically expanding preserved tissues embedded in a swellable hydrogel. Biomolecules of interest are covalently linked to a polymer matrix, which then isotopically expands at least 100-fold in size in pure water after mechanical homogenization of the tissue-gel. The sample can then be investigated with nanoscale precision using a conventional diffraction-limited microscope. The protocol described here is a variant of ExM that uses regents and equipment found in a typical biology laboratory and has been optimized for imaging proteins in expanded brain tissues.

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