Expansion Microscopy for Beginners: Visualizing Microtubules in Expanded Cultured HeLa Cells
Name
cpns.96.pdf
Description
Published version
Size
1.39 MB
Format
Adobe PDF
Checksum (MD5)
5fb02b0e33e16cda5a829cd01319d005
Author(s)
Zhang, Chi
Kang, Jeong Seuk
Asano, Shoh M
Gao, Ruixuan
Boyden, Edward S
Date Issued
2020
Journal
Current Protocols in Neuroscience
Publisher
Wiley
Version
Final published version
Abstract
© 2020 The Authors. Expansion microscopy (ExM) is a technique that physically expands preserved cells and tissues before microscope imaging, so that conventional diffraction-limited microscopes can perform nanoscale-resolution imaging. In ExM, biomolecules or their markers are linked to a dense, swellable gel network synthesized throughout a specimen. Mechanical homogenization of the sample (e.g., by protease digestion) and the addition of water enable isotropic swelling of the gel, so that the relative positions of biomolecules are preserved. We previously presented ExM protocols for analyzing proteins and RNAs in cells and tissues. Here we describe a cookbook-style ExM protocol for expanding cultured HeLa cells with immunostained microtubules, aimed to help newcomers familiarize themselves with the experimental setups and skills required to successfully perform ExM. Our aim is to help beginners, or students in a wet-lab classroom setting, learn all the key steps of ExM. © 2020 The Authors.
MIT Department
Massachusetts Institute of Technology. Media Laboratory
McGovern Institute for Brain Research at MIT
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1002/CPNS.96
