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.
The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-8719-1_19
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Change history
24 October 2018
This book was inadvertently published with the incorrect title as Clathrin-Mediated Endoytosis: Methods and Protocols. This has now been corrected throughout the book to Clathrin-Mediated Endocytosis: Methods and Protocols.
References
Hell SW, Wichmann J (1994) Breaking the diffraction resolution limit by stimulated emission: stimulated-emission-depletion fluorescence microscopy. Opt Lett 19(11):780–782
Schermelleh L, Heintzmann R, Leonhardt H (2010) A guide to super-resolution fluorescence microscopy. J Cell Biol 190(2):165–175
Tonnesen J, Nadrigny F, Willig KI, Wedlich-Soldner R, Nagerl UV (2011) Two-color STED microscopy of living synapses using a single laser-beam pair. Biophys J 101(10):2545–2552
Lukinavicius G, Umezawa K, Olivier N, Honigmann A, Yang G, Plass T, Mueller V, Reymond L, Correa IR Jr, Luo ZG, Schultz C, Lemke EA, Heppenstall P, Eggeling C, Manley S, Johnsson K (2013) A near-infrared fluorophore for live-cell super-resolution microscopy of cellular proteins. Nat Chem 5(2):132–139
Bottanelli F, Kromann EB, Allgeyer ES, Erdmann RS, Wood Baguley S, Sirinakis G, Schepartz A, Baddeley D, Toomre DK, Rothman JE, Bewersdorf J (2016) Two-colour live-cell nanoscale imaging of intracellular targets. Nat Commun 7:10778
Abramoff MD, Magalhaes PJ, Ram SJ (2004) Image processing with ImageJ. Biophoton Int 11(7):36–42
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.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bottanelli, F., Schroeder, L. (2018). Stimulated Emission Depletion (STED) Imaging of Clathrin-Mediated Endocytosis in Living Cells. In: Swan, L. (eds) Clathrin-Mediated Endocytosis. Methods in Molecular Biology, vol 1847. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8719-1_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8719-1_14
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8717-7
Online ISBN: 978-1-4939-8719-1
eBook Packages: Springer Protocols