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

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Riccardo Maraspini - , Max-Planck-Institut für molekulare Zellbiologie und Genetik (Autor:in)
  • Oliver Beutel - , Max-Planck-Institut für molekulare Zellbiologie und Genetik (Autor:in)
  • Alf Honigmann - , Max-Planck-Institut für molekulare Zellbiologie und Genetik (Autor:in)

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

OriginalspracheEnglisch
Seiten (von - bis)188-197
Seitenumfang10
FachzeitschriftMethods
Jahrgang140-141
PublikationsstatusVeröffentlicht - 1 Mai 2018
Peer-Review-StatusJa
Extern publiziertJa

Externe IDs

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

Schlagworte

Schlagwörter

  • 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