TY - JOUR
T1 - Expansion microscopy of c. Elegans
AU - Yu, Chih Chieh
AU - Barry, Nicholas C.
AU - Wassie, Asmamaw T.
AU - Sinha, Anubhav
AU - Bhattacharya, Abhishek
AU - Asano, Shoh
AU - Zhang, Chi
AU - Chen, Fei
AU - Hobert, Oliver
AU - Goodman, Miriam B.
AU - Haspel, Gal
AU - Boyden, Edward S.
N1 - Funding Information:
The authors thank Steven W Flavell for sharing the transgenic strain CX16682. The authors thank Steven W Flavell, Piali Sengupta, Zainab Tanvir, Kiryl Piatkevich, Grace Huynh, Adam Marblestone, Anna Kazatskaya and Inna Nechipurenko for helpful discussions. C-CY acknowledges the McGovern Institute for Brain Research at MIT for the Friends of the McGovern Fellowship. GH acknowledges the New Jersey Institute of Technology for the Faculty Seed Grant. ESB acknowledges, for funding, Lisa Yang, John Doerr, the Open Philanthropy Project, NIH 1R01EB024261, NSF Grant 1734870, the HHMI-Simons Faculty Scholars Program, US Army Research Laboratory and the US Army Research Office under contract/grant number W911NF1510548, NIH 1R01MH103910, NIH 1R01MH114031, NIH 1R01MH110932, IARPA D16PC00008, NIH 2R01DA029639, and NIH 1RM1HG008525.
Publisher Copyright:
© Yu et al.
PY - 2020/5
Y1 - 2020/5
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85084879905&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85084879905&partnerID=8YFLogxK
U2 - 10.7554/eLife.46249
DO - 10.7554/eLife.46249
M3 - Article
C2 - 32356725
AN - SCOPUS:85084879905
SN - 2050-084X
VL - 9
SP - 1
EP - 78
JO - eLife
JF - eLife
M1 - e46249
ER -