Dual-Color and 3D Super-Resolution Microscopy of Multi-protein Assemblies

  • Philipp Hoess
  • Markus Mund
  • Manuel Reitberger
  • Jonas Ries
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1764)

Abstract

Breaking the resolution limit of conventional microscopy by super-resolution microscopy (SRM) led to many new biological insights into protein assemblies at the nanoscale. Here we provide detailed protocols for single-molecule localization microscopy (SMLM) to image the structure of a protein complex. As examples, we show how to acquire single- and dual-color super-resolution images of the nuclear pore complex (NPC) and dual-color 3D data on actin and paxillin in focal adhesions.

Key words

Super-resolution microscopy Single-molecule localization microscopy PALM STORM Nuclear pore complex Focal adhesions Photoswitchable fluorescent protein 

Notes

Acknowledgments

We thank Ulf Matti for experimental assistance and Edward Lemke for the Flp-In™ T-Rex™ 293 cell line. This work was supported by EMBL International PhD Programme fellowships (P.H. and M.M.).

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Copyright information

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

Authors and Affiliations

  • Philipp Hoess
    • 1
  • Markus Mund
    • 1
  • Manuel Reitberger
    • 1
  • Jonas Ries
    • 1
  1. 1.Cell Biology and Biophysics UnitEuropean Molecular Biology LaboratoryHeidelbergGermany

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