Imaging fluorescence correlation spectroscopy studies lipid membrane dynamics and organization: Applications to the investigation of lipid bilayers, cell membranes, and membrane active peptides

Abstract

The investigation of the spatio-temporal dynamics and organization of plasma membrane requires advanced biophysical methods which can track its physical properties. Imaging FCS is a single molecule sensitive technique that can provide maps of molecular dynamics and concentration of a large membrane area under physiological conditions. In the first part of the thesis, the instrumental factors affecting the accuracy and precision of the Imaging FCS data are optimized and the protocol for the data evaluation is developed with extensive calibration experiments on an array of model membranes. This is followed by the integration of the analytical tool called FCS diffusion law to the Imaging FCS framework to study sub-resolution membrane organization. The second part describes the applications of the Imaging FCS to study membrane physical chemistry and peptide-membrane interaction. Imaging FCS proves to be a valuable tool with sufficient spatial and temporal resolution to study membrane processes.