Interests and limits of continuous excitation photo-physical methods applied to cellular rheology

Abstract

It is possible to characterize the cohesion of the lipidic zones in cells or in membranes by using molecular emission spectroscopy techniques such as: fluorescence polarization, quenching reactions, intramolecular reactions.

However, applying these techniques in biology can be improved by the use of pulsed excitations leading to the time-resolved evolutions of some properties of the probe, which can themselves be a function of their environment. The first and the last techniques, based on the use of diphenylhexatriene and dipyrenylpyopane respectively are used in practice by steady state excitation. In the case of fluorescence polarization, it can be shown that the method can be applied, within certain limits, which have to be precised when studying the modifications in the membranes. This is also partly true for methods based on the use of intramolecular reactions. However these continuous excitation techniques usually only lead to a global result which is a function of several spectroscopic parameters and of the “cohesion” of the membrane. Then it can be shown that to go beyond this step of semi-quantitative analysis, it is necessary to use pulsed or modulated excitation methods. Moreover this more sophisticated technical approach implies a new theoretical effort to understand the molecular dynamics of the membranes.