Triple-Color Super-Resolution Imaging of Live Cells: Resolving Submicroscopic Receptor Organization in the Plasma Membrane

Authors


  • We thank Gabriele Hikade and Hella Kenneweg for technical support, Roland Wedlich-Söldner (MPI Martinsried) for providing a plasmid coding for Lifeact, Volker Haucke (Freie Universität Berlin) for providing a plasmid coding for clathrin light chain, and Vladislav Verkhusha (Albert Einstein College of Medicine, New York) for a plasmid coding for PAtagRFP. This project was supported by funding from the DFG (PI405-5 and SFB 944), the European Community’s Seventh Framework Programme (FP7/2007-2013) under grant agreement number 223608 (IFNaction), and the National Institute of General Medical Sciences (GM094713).

Abstract

original image

In living color: Efficient intracellular covalent labeling of proteins with a photoswitchable dye using the HaloTag for dSTORM super-resolution imaging in live cells is described. The dynamics of cellular nanostructures at the plasma membrane were monitored with a time resolution of a few seconds. In combination with dual-color FPALM imaging, submicroscopic receptor organization within the context of the membrane skeleton was resolved (see figure).

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