Circle scanning STED fluorescence correlation spectroscopy to quantify membrane dynamics and compartmentalization

Research output: Contribution to journalResearch articleContributedpeer-review

Contributors

  • Riccardo Maraspini - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Oliver Beutel - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)
  • Alf Honigmann - , Max Planck Institute of Molecular Cell Biology and Genetics (Author)

Abstract

Quantifying molecular dynamics of cell membrane constituents is required to understand organization and function of biological membranes. Because of its complex structure unambiguous interpretation of molecular membrane dynamics requires high spatial and temporal resolution measurements. In this paper, we provide a comprehensive description of circle scanning fluorescence correlation spectroscopy and its combination with stimulated emission depletion microscopy (CS-STED-FCS). This method allows quantification of sub-diffusion processes and direct mapping of heterogeneities in membranes with high spatiotemporal resolution. We show how to use model membranes to calibrate and test the technique and how to apply it in the context of living cells to quantify membrane dynamics with high spatiotemporal resolution and good statistics.

Details

Original languageEnglish
Pages (from-to)188-197
Number of pages10
JournalMethods
Volume140-141
Publication statusPublished - 1 May 2018
Peer-reviewedYes
Externally publishedYes

External IDs

Scopus 85038885705
ORCID /0000-0003-0475-3790/work/155291294

Keywords

Keywords

  • Animals, Calibration, Cell Line, Cell Membrane/metabolism, Diffusion, Fluorescent Dyes/chemistry, Humans, Image Processing, Computer-Assisted/methods, Intravital Microscopy/instrumentation, Lipid Bilayers/metabolism, Membrane Proteins/metabolism, Microscopy, Fluorescence/instrumentation, Molecular Dynamics Simulation, Spectrometry, Fluorescence/instrumentation