Fluorescence lifetime imaging provides enhanced contrast when imaging the phase-sensitive dye di-4-ANEPPDHQ in model membranes and live cells

Biophys J. 2006 Jun 1;90(11):L80-2. doi: 10.1529/biophysj.106.084673. Epub 2006 Apr 14.


We apply fluorescence lifetime imaging to the membrane phase-sensing dye di-4-ANEPPDHQ in model membranes and live cells. We show that the 1700 ps lifetime shift between liquid-disordered and liquid-ordered phases offers greater contrast than the 60 nm spectral shift previously reported. Detection of cholesterol-rich membrane microdomains is confirmed by observation of the temperature dependence of membrane order and by cholesterol depletion using methyl-beta-cyclodextrin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Cell Membrane / chemistry*
  • Cell Membrane / ultrastructure
  • Cholesterol / chemistry
  • Fluorescent Dyes / chemistry*
  • Humans
  • Image Enhancement
  • Lipid Bilayers / chemistry*
  • Membrane Microdomains / chemistry*
  • Membrane Microdomains / ultrastructure
  • Temperature
  • beta-Cyclodextrins / chemistry


  • Fluorescent Dyes
  • Lipid Bilayers
  • beta-Cyclodextrins
  • methyl-beta-cyclodextrin
  • Cholesterol