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Stimulated Emission Depletion (STED) Imaging of Clathrin-Mediated Endocytosis in Living Cells

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1847)

Abstract

The recent development of probes and labeling strategies for multicolor super-resolution imaging in living cells allows cell biologists to follow cellular processes with unprecedented details. Here we describe how to image endocytic events at the plasma membrane of living cells using commercial (Leica, Abberior Instruments) or custom built STED microscopes.

Key words

STED Super-resolution imaging Clathrin Transferrin receptor Endocytosis Halo tag SNAP tag 

Notes

Acknowledgments

We thank Stephanie Wood Baguley for generating the plasmids used in this protocol. We thank Dr. Joerg Bewersdorf, Dr. Manuel Jütte, Dr. Edward Allgeyer, and Mark Lessard for critically reading the manuscript. This project was supported by the Wellcome Trust (095927/A/11/Z, 092096) and the G. Harold & Leila Y. Mathers Charitable Foundation.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Cell BiologyYale School of MedicineNew HavenUSA

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