A Förster resonance energy transfer (FRET) approach for enhancing fluorescence contrast in phase-separated membranes

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DOIResolve DOI: http://doi.org/10.1139/V10-144
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TypeArticle
Journal titleCanadian Journal of Chemistry
Volume89
Pages423432; # of pages: 10
Subjectfluorescence microscopy; lipid bilayers; vesicles; ceramide; bicouches de lipide; vésicules; céramide
AbstractThe partitioning of the dye-labeled lipid probe, NBD-DHPE (1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (ammonium salt)), was examined by fluorescence microscopy in phase-separated lipid bilayers with mixtures of coexisting liquid-ordered and fluid phases. This probe shows slightly higher fluorescence intensity in the ordered domains but undergoes a contrast reversal to give a more strongly fluorescent fluid phase in the presence of>0.2% Texas red-DHPE (TR-DHPE). The change in contrast is shown to result from Förster resonance energy transfer between the NBD donor and TR acceptor in the fluid phase, which has a TR concentration that is approximately 3 times higher than in the domains. An alternate approach using a nitroxide-substituted lipid that partitions into the fluid phase as a quencher, was also examined as a means to enhance the contrast; however, the quencher modified the behaviour of the bilayer. The energy transfer method for enhancing the contrast between ordered and fluid phases was used to examine the morphology of enzyme-treated bilayers.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Steacie Institute for Molecular Sciences
Peer reviewedYes
NPARC number17652985
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Record identifier42f6e266-5262-4592-9061-0bca57efa47e
Record created2011-03-31
Record modified2016-05-09
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