Fluorescence lifetime imaging provides enhanced contrast when imaging the phase-sensitive dye di-4-ANEPPDHQ in model membranes and live cells.
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.
Owen
Dylan M
DM
Chemical Biology Centre, Department of Physics, Imperial College London, London, United Kingdom. dylan.owen@imperial.ac.uk
Lanigan
Peter M P
PM
Dunsby
Christopher
C
Munro
Ian
I
Grant
David
D
Neil
Mark A A
MA
French
Paul M W
PM
Magee
Anthony I
AI
D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure; D002784:Cholesterol / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007089:Image Enhancement; D008051:Lipid Bilayers / Q000737:chemistry*; D021962:Membrane Microdomains / Q000737:chemistry* / Q000648:ultrastructure; D013696:Temperature; D047392:beta-Cyclodextrins / Q000737:chemistry
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D002784:Cholesterol