Cholesterol-enriched lipid domains can be visualized by di-4-ANEPPDHQ with linear and nonlinear optics

Biophys J. 2005 Jul;89(1):L04-6. doi: 10.1529/biophysj.105.064816. Epub 2005 May 6.


We present a membrane-staining dye, di-4-ANEPPDHQ, which differentiates liquid-ordered phases from liquid-disordered phases coexisting in model membranes under both linear and nonlinear microscopies. The dye's fluorescence emission spectrum is blue-shifted 60 nm in liquid-ordered phases compared with liquid-disordered phases, and shows strong second harmonic generation in the liquid-disordered phase compared with the liquid-ordered phase. The ease of staining and the ability of this single dye to detect both phases, should lead to broad applications in biophysical studies of lipid domains in model membranes and cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biophysics / instrumentation
  • Biophysics / methods*
  • Cell Membrane / metabolism
  • Cholesterol / chemistry*
  • Fluorescent Dyes / pharmacology*
  • Lipid Bilayers / chemistry
  • Lipids / chemistry*
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Oscillometry
  • Protein Structure, Tertiary
  • Spectrometry, Fluorescence
  • Spectrophotometry
  • Water / chemistry


  • Fluorescent Dyes
  • Lipid Bilayers
  • Lipids
  • Water
  • Cholesterol