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Observing the Assembly of Protein Complexes in Living Eukaryotic Cells in Super-Resolution Using refSOFI

  • Fabian Hertel
  • Gary C. H. Mo
  • Peter Dedecker
  • Jin Zhang
Part of the Methods in Molecular Biology book series (MIMB, volume 1764)

Abstract

Few approaches are currently available that allow the detection of protein-protein interactions (PPIs) in super-resolution, and the observation of the assembly of protein complexes in living cells has been particularly challenging. We developed reconstituted fluorescence-based stochastic optical fluctuation imaging (refSOFI), which is based on bimolecular fluorescence complementation (BiFC) and SOFI, allowing us to detect protein complex assembly 30 min after the induction of complex formation. Here we describe how to use refSOFI to map the assembly of two proteins of interest into a complex within living cells at super-resolution.

Key words

Super-resolution imaging Stochastic optical fluctuation imaging (SOFI) Protein complexes Protein-protein interactions Bimolecular fluorescence complementation (BiFC) 

Notes

Acknowledgment

This work was supported by NIH (R35 CA197622, R01 DK073368, R01 GM111665, and R01 MH111516 to J.Z.).

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Copyright information

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

Authors and Affiliations

  • Fabian Hertel
    • 1
  • Gary C. H. Mo
    • 2
  • Peter Dedecker
    • 1
  • Jin Zhang
    • 2
  1. 1.Department of ChemistryUniversity of LeuvenHeverleeBelgium
  2. 2.Department of PharmacologyUniversity of California at San DiegoLa JollaUSA

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