IMAGING FLUORESCENCE CORRELATION SPECTROSCOPY: THEORY, SIMULATIONS AND APPLICATIONS TO PROBE LIPID-MEMBRANE DYNAMICS

Abstract

The cell membrane is organized in to different structures and it has been difficult to visualize these structures since they are believed to possess sizes below the optical resolution limit. Hence the development of new tools probing the biophysical properties of membranes is necessary. Imaging FCS is one such tool which allows the measurement of mobility at contiguous locations on cell membranes of live cells by analyzing the autocorrelation functions. In this thesis, Imaging FCS is being extended to Imaging FCCS enabling one to calculate cross-correlations and to extract parameters from the same. The next part discusses methods to characterize organization of membranes. The last part describes the study of membrane proteins and anti-microbials carried out using Imaging FCS. Unlike single point FCS which yields only mobility, imaging FCS provides mobility and heterogeneity and proved to be a valuable biophysical tool to characterize the dynamics and organization of cell membranes.