Expansion microscopy of C. elegans
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 C. elegans is challenged by its cuticle, which is stiff and impermeable to antibodies. Here we present a strategy, expansion of C. elegans (ExCel), to expand fixed, intact C. elegans. 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.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- May 01, 2020
- Source ID
- 10.7554/elife.46249
Entities
People
- Abhishek Bhattacharya
- Anubhav Sinha
- Asmamaw T. Wassie
- Chi Zhang
- Chih-Chieh Jay Yu
- Edward Boyden
- Fei Chen
- Gal Haspel
- Lisa Yang
- Miriam B. Goodman
- Nicholas C Barry
- Oliver Hobert
- Shoh M Asano
Organizations
- Army Research Office
- Broad Institute
- Columbia University
- Howard Hughes Medical Institute
- Intelligence Advanced Research Projects Activity
- Massachusetts Institute of Technology
- McGovern Institute for Brain Research
- National Institutes of Health
- National Science Foundation
- New Jersey Institute of Technology
- Open Philanthropy Project
- Stanford University
- United States Army Research Laboratory