Nonlinear Optical Properties of Fluorescent Dyes Allow for Accurate Determination of Their Molecular Orientations in Phospholipid Membranes

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• 作者：艩t臎p谩n Timr ; Ji艡铆 Brabec ; Alexey Bondar ; Tom谩拧 Ryba ; Milo拧 沤elezn媒 ; Josef Lazar ; Pavel Jungwirth
• 刊名：Journal of Physical Chemistry B
• 出版年：2015
• 出版时间：July 30, 2015
• 年：2015
• 卷：119
• 期：30
• 页码：9706-9716
• 全文大小：735K
• ISSN：1520-5207

文摘

Several methods based on single- and two-photon fluorescence detected linear dichroism have recently been used to determine the orientational distributions of fluorescent dyes in lipid membranes. However, these determinations relied on simplified descriptions of nonlinear anisotropic properties of the dye molecules, using a transition dipole-moment-like vector instead of an absorptivity tensor. To investigate the validity of the vector approximation, we have now carried out a combination of computer simulations and polarization microscopy experiments on two representative fluorescent dyes (DiI and F2N12S) embedded in aqueous phosphatidylcholine bilayers. Our results indicate that a simplified vector-like treatment of the two-photon transition tensor is applicable for molecular geometries sampled in the membrane at ambient conditions. Furthermore, our results allow evaluation of several distinct polarization microscopy techniques. In combination, our results point to a robust and accurate experimental and computational treatment of orientational distributions of DiI, F2N12S, and related dyes (including Cy3, Cy5, and others), with implications to monitoring physiologically relevant processes in cellular membranes in a novel way.