Optimized time-gated generalized polarization imaging of Laurdan and di-4-ANEPPDHQ for membrane order image contrast enhancement.
The membrane dyes Laurdan and di-4-ANEPPDHQ can be used to image membrane order due to a spectral blue-shift in the fluorescence emission between the liquid-ordered and liquid-disordered phases. These images typically take the form of a normalized intensity ratio image known as a generalized polarization (GP) plot. Here, we exploit the known excited state photophysics and time-resolved data acquisition via time-correlated single-photon counting (TCSPC) to demonstrate GP contrast enhancement for these two probes of 7 and 31%, respectively. This improvement in image contrast enhancement will be invaluable when studying the role of lipid rafts in fixed and live cell systems.
Owen
Dylan M
DM
Centre for Vascular Research, University of New South Wales, Sydney, Australia. dylan.owen@unsw.edu.au
Gaus
Katharina
K
D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure*; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007848:Laurates / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D011726:Pyridinium Compounds / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*; D013997:Time Factors
D007848:Laurates; D011726:Pyridinium Compounds; C541550:di-4-ANEPPDHQ; D015081:2-Naphthylamine; C065580:laurdan