Modern Research and Educational Topics in Microscopy

SANCHEZ, S. A.; TRICERRI, M. A.; GUNTHER, G.; GRATTON, E.

Microscopy Book Series

Año: 2007;

Laurdan is a fluorescent molecule that detects changes in membrane phase properties through its sensitivity to the polarity of the environment in the bilayer. Polarity changes are shown by shifts in the Laurdan emission spectrum, which are quantified by calculating the generalized polarization (GP). This technique was originally developed to be used in a cuvette fluorometer. With the development of two-photon microscopy, Laurdan GP has evolved into a technique capable of spatially resolve micro-domains of different solvent penetration. We discuss here the basic concepts, instrumentation and experimental considerations when transferring the GP technique from the cuvette to the microscope. We also discuss examples of Laurdan GP in membrane model systems using both cuvette and microscope approaches to compare the two methodologies.