Expansion microscopy of c. Elegans
Document Type
Article
Publication Date
5-1-2020
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.
Identifier
85084879905 (Scopus)
Publication Title
Elife
External Full Text Location
https://doi.org/10.7554/eLife.46249
e-ISSN
2050084X
PubMed ID
32356725
First Page
1
Last Page
78
Volume
9
Grant
T32EB001680
Fund Ref
Army Research Laboratory
Recommended Citation
Yu, Chih Chieh; Barry, Nicholas C.; Wassie, Asmamaw T.; Sinha, Anubhav; Bhattacharya, Abhishek; Asano, Shoh; Zhang, Chi; Chen, Fei; Hobert, Oliver; Goodman, Miriam B.; Haspel, Gal; and Boyden, Edward S., "Expansion microscopy of c. Elegans" (2020). Faculty Publications. 5344.
https://digitalcommons.njit.edu/fac_pubs/5344
