Imaging Fluorescent Nuclear Pore Complex Proteins in C. elegans

Courtney Lancaster; Giulia Zavagno; James Groombridge; Adelaide Raimundo; David Weinkove; Tim Hawkins; Joanne Robson; Martin W Goldberg

doi:10.1007/978-1-0716-2337-4_24

Imaging Fluorescent Nuclear Pore Complex Proteins in C. elegans

Methods Mol Biol. 2022:2502:373-393. doi: 10.1007/978-1-0716-2337-4_24.

Authors

Courtney Lancaster¹, Giulia Zavagno², James Groombridge², Adelaide Raimundo², David Weinkove², Tim Hawkins², Joanne Robson², Martin W Goldberg³

Affiliations

¹ MRC Laboratory for Molecular Cell Biology, University College London, London, UK.
² Department of Biosciences, Durham University, Durham, UK.
³ Department of Biosciences, Durham University, Durham, UK. m.w.goldberg@durham.ac.uk.

PMID: 35412251
DOI: 10.1007/978-1-0716-2337-4_24

Abstract

C. elegans is a well-characterized and relatively simple model organism, making it attractive for studying nuclear pore complex proteins in cell and developmental biology. C. elegans is transparent and highly amendable to genetic manipulation. Therefore, it is possible to generate fluorescently tagged proteins and combine this with various light microscopy techniques to study protein behavior in space and time. Here, we provide protocols to prepare both fixed and live C. elegans for confocal and light sheet microscopy. This enables the analysis of nuclear pore complex proteins from embryonic stages to the aging adult.

Keywords: 3D-SIM; C. elegans; Confocal; Fluorescence microscopy; Light sheet; Nuclear pore complex; Nucleus; Superresolution; fluorescent protein.

MeSH terms

Animals
Caenorhabditis elegans Proteins* / genetics
Caenorhabditis elegans Proteins* / metabolism
Caenorhabditis elegans* / genetics
Caenorhabditis elegans* / metabolism
Microscopy, Fluorescence / methods
Nuclear Pore Complex Proteins* / metabolism

Substances

Caenorhabditis elegans Proteins
Nuclear Pore Complex Proteins