Using FM Dyes to Monitor Clathrin-Mediated Endocytosis in Primary Neuronal Culture

Michael A Cousin; Sarah L Gordon; Karen J Smillie

doi:10.1007/978-1-4939-8719-1_18

Using FM Dyes to Monitor Clathrin-Mediated Endocytosis in Primary Neuronal Culture

Methods Mol Biol. 2018:1847:239-249. doi: 10.1007/978-1-4939-8719-1_18.

Authors

Michael A Cousin¹, Sarah L Gordon², Karen J Smillie³

Affiliations

¹ Centre for Discovery Brain Sciences, Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK.
² Florey Institute of Neuroscience and Mental Health, The University of Melbourne, Parkville, VIC, Australia.
³ Centre for Discovery Brain Sciences, Edinburgh Medical School: Biomedical Sciences, University of Edinburgh, Edinburgh, UK. K.Smillie@ed.ac.uk.

PMID: 30129022
DOI: 10.1007/978-1-4939-8719-1_18

Abstract

This protocol utilizes lipophilic FM dyes to monitor membrane recycling in real time. FM dyes are virtually nonfluorescent in solution but when membrane bound are intensely fluorescent, combined with the flexibility of different emission wavelengths make these dyes an excellent choice for investigating clathrin-mediated endocytosis, among other membrane trafficking and recycling pathways.

Keywords: Clathrin-mediated endocytosis; Epifluorescence; FM dyes; Live cell imaging; Primary neuronal culture.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cells, Cultured
Clathrin / metabolism
Endocytosis*
Fluorescent Dyes / chemistry*
Microscopy, Fluorescence / methods*
Neurons / metabolism

Substances

Clathrin
Fluorescent Dyes

Grants and funding

G1002117/Medical Research Council/United Kingdom