Expansion microscopy of C. Elegans

Elife
Authors
Abstract

We recently developed expansion microscopy (ExM), which achieves nanoscale-precise imaging of specimens at ~70 nm resolution (with ~4.5x linear expansion) by isotropic swelling of chemically processed, hydrogel-embedded tissue. ExM of is challenged by its cuticle, which is stiff and impermeable to antibodies. Here we present a strategy, expansion of (ExCel), to expand fixed, intact . ExCel enables simultaneous readout of fluorescent proteins, RNA, DNA location, and anatomical structures at resolutions of ~65-75 nm (3.3-3.8x linear expansion). We also developed epitope-preserving ExCel, which enables imaging of endogenous proteins stained by antibodies, and iterative ExCel, which enables imaging of fluorescent proteins after 20x linear expansion. We demonstrate the utility of the ExCel toolbox for mapping synaptic proteins, for identifying previously unreported proteins at cell junctions, and for gene expression analysis in multiple individual neurons of the same animal.

Year of Publication
2020
Journal
Elife
Volume
9
Date Published
2020 May 01
ISSN
2050-084X
DOI
10.7554/eLife.46249
PubMed ID
32356725
PubMed Central ID
PMC7195193
Links
Grant list
1R01EB024261 / NH / NIH HHS / United States
1R01MH110932 / NH / NIH HHS / United States
1734870 / National Science Foundation
Faculty Seed Grant / New Jersey Institute of Technology
D16PC00008 / Intelligence Advanced Research Projects Activity
HHMI-Simons Faculty Scholars Program / HHMI / Howard Hughes Medical Institute / United States
R01 MH110932 / MH / NIMH NIH HHS / United States
Friends of the McGovern Fellowship / McGovern Institute for Brain Research at MIT
1R01MH103910 / NH / NIH HHS / United States
W911NF1510548 / U.S. Army Research Laboratory
W911NF1510548 / Army Research Office
R01 DA029639 / DA / NIDA NIH HHS / United States
R01 MH114031 / MH / NIMH NIH HHS / United States
1R01MH114031 / NH / NIH HHS / United States
1RM1HG008525 / NH / NIH HHS / United States
RM1 HG008525 / HG / NHGRI NIH HHS / United States
R01 MH103910 / MH / NIMH NIH HHS / United States
2R01DA029639 / NH / NIH HHS / United States
R01 EB024261 / EB / NIBIB NIH HHS / United States