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Originally published as Biophys J. BioFAST on April 14, 2006.
doi:10.1529/biophysj.106.084673
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Biophysical Journal 90:L80-L82 (2006)
© 2006 The Biophysical Society

Fluorescence Lifetime Imaging Provides Enhanced Contrast when Imaging the Phase-Sensitive Dye di-4-ANEPPDHQ in Model Membranes and Live Cells

Dylan M. Owen *, Peter M. P. Lanigan {dagger}, Christopher Dunsby {dagger}, Ian Munro {dagger}, David Grant *, Mark A. A. Neil {dagger}, Paul M. W. French {dagger} and Anthony I. Magee {ddagger}

* Chemical Biology Centre, {dagger} Department of Physics, and {ddagger} Division of Biomedical Sciences, Imperial College London, London, United Kingdom

Correspondence: Address reprint requests and inquiries to Dylan M. Owen, E-mail: dylan.owen{at}imperial.ac.uk.

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-ß-cyclodextrin.




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