Cholesterol-enriched lipid domains can be visualized by di-4-ANEPPDHQ with linear and nonlinear optics.
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.
Jin
Lei
L
Department of Cell Biology and Center for Cell Analysis and Modeling, University of Connecticut Health Center, Farmington, Connecticut 06030-1507, USA.
Millard
Andrew C
AC
Wuskell
Joseph P
JP
Clark
Heather A
HA
Loew
Leslie M
LM
D001703:Biophysics / Q000295:instrumentation / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism; D002784:Cholesterol / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000494:pharmacology*; D008051:Lipid Bilayers / Q000737:chemistry; D008055:Lipids / Q000737:chemistry*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009991:Oscillometry; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry; D014867:Water / Q000737:chemistry
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008055:Lipids; D014867:Water; D002784:Cholesterol