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Biophys. J. BioFAST: First Published April 14, 2006. doi:10.1529/biophysj.106.084673
© 2006 by the Biophysical Society.

A more recent version of this article appeared on June 1, 2006.
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BIOPHYSICAL LETTERS

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

Dylan M. Owen 1*, Peter M. P. Lanigan 1, Christopher Dunsby 1, Ian Munro 1, David M. Grant 1, Mark A. A. Neil 1, Paul M. W. French 1 and Anthony I. Magee 1

1 Imperial College London

* To whom correspondence should be addressed. E-mail: dylan.owen{at}imperial.ac.uk.

Submitted on March 8, 2006
Revised on March 24, 2006
Accepted on 7 April 2006


  Abstract
We apply fluorescence lifetime imaging (FLIM) to the membrane phase-sensing dye di-4-ANEPPDHQ in model membranes and live cells. We show that the 1700ps lifetime shift between liquid-disordered and liquid-ordered phases offers greater contrast than the 60nm 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.

Key Words: FLIM, fluorescence lifetime imaging, ordered membrane, rafts




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Copyright © 2006 by the Biophysical Society.