Super-resolution optical microscopy of lipid plasma membrane dynamics

Essays Biochem. 2015:57:69-80. doi: 10.1042/bse0570069.

Abstract

Plasma membrane dynamics are an important ruler of cellular activity, particularly through the interaction and diffusion dynamics of membrane-embedded proteins and lipids. FCS (fluorescence correlation spectroscopy) on an optical (confocal) microscope is a popular tool for investigating such dynamics. Unfortunately, its full applicability is constrained by the limited spatial resolution of a conventional optical microscope. The present chapter depicts the combination of optical super-resolution STED (stimulated emission depletion) microscopy with FCS, and why it is an important tool for investigating molecular membrane dynamics in living cells. Compared with conventional FCS, the STED-FCS approach demonstrates an improved possibility to distinguish free from anomalous molecular diffusion, and thus to give new insights into lipid-protein interactions and the traditional lipid 'raft' theory.

MeSH terms

  • Diffusion
  • Fluorescent Dyes
  • Lipid Bilayers / chemistry*
  • Lipid Bilayers / metabolism
  • Membrane Microdomains / chemistry
  • Membrane Microdomains / ultrastructure*
  • Membrane Proteins / chemistry
  • Membrane Proteins / ultrastructure
  • Microscopy, Confocal / instrumentation*
  • Microscopy, Confocal / methods
  • Phosphatidylcholines / chemistry
  • Phosphatidylethanolamines / chemistry
  • Spectrometry, Fluorescence / instrumentation*
  • Spectrometry, Fluorescence / methods
  • Sphingomyelins / chemistry

Substances

  • Fluorescent Dyes
  • Lipid Bilayers
  • Membrane Proteins
  • Phosphatidylcholines
  • Phosphatidylethanolamines
  • Sphingomyelins
  • phosphatidylethanolamine
  • 1,2-oleoylphosphatidylcholine