The Use of Fluorescence Lifetimes for Membrane Organizational Studies

The fluorescence lifetime is sensitive to the environment of the fluorophore, and in membranes this usually means the surrounding fatty acyl chains or the membrane protein interfacial region (see summary in Table 5.3). Generally, the lifetime of membrane-bound fluorophores is rather less sensitive to the types of subtle alterations which are encountered in membranes as compared to the fluorescence anisotropy parameters. The gel-to-liquid crystalline phase transition is a notable exception where most fluorophores show an alteration in lifetime properties. Although, again, the anisotropy (see below) is the most sensitive parameter in this regard, the fluorescence lifetime has been used with considerable success in the study of phase transitions and lateral phase separations. Fluorophores used to yield information on the 

The use of DPH lifetimes for the analysis of phase separations and membrane properties has been described for mode) systems.n fl) In the case of both parinaric acids and DPH, one of the motivations for examining phase separation in a model lipid bilayer is the possibility that phase separations might be detectable in natural membranes. However, this technique has not been able to satisfactorily resolve lateral phase separations in natural membranes, either because they do not exist or because they are much more complex and even possibly transient in nature. Alternatively, it could be argued that if a probe could be found with the characteristics of trans-parinaric acid but perhaps with an even greater phase partitioning ability, then this approach might be reevaluated. 

Another factor affecting the lifetime of a membrane fluorophore probe is its proximity to the surface. The lifetimes of the DPH, DPH-phosphatidyl-choline (DPH-PC), and trimethylammonium-DPH (TMA-DPH) probes decrease in the order DPH DPH-PC TMA-DPH, as the probe locates nearer to the surface of the lipid bilayer.(7) The same is found for the anthroyl-stearate probes.(8) More recently, it has been shown that with TMA-DPH, the lifetime appears to be fairly sensitive to the differences in lipid bilayer packing induced by differing degrees of unsaturation in the phospholipid fatty acyl chains.(9) This aspect of the use of TMA-DPH and possibly other probes remains to be further exploited. 

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