Characterizing lateral diffusion of EGFR using advanced fluorescence spatial correlation functions

Epidermal growth factor receptor (EGFR) has various functionalities in the cell cycle and also plays an important role in cellular transport. EGFR is found to be localized on the cell membrane before its activation. Although, there are numerous studies that focus on the transport and signaling mechanism of EGFR mediated cellular transport, only few studies have been done in the area characterizing its lateral diffusion. In our work, we focus on characterizing EGFR lateral diffusion prior to and after activation of the receptor by epidermal growth factor using Total Internal Fluorescence Reflection Microscopy (TIRF). TIRF microscopy uses camera based data collection and novel pixel based correlation methods enable us to analyze and study a diffusing population. One can record whole cell, and analyze and decipher the predominant diffusion mechanisms presented in the cell using the techniques like pair-correlation function, image mean square displacement, number and brightness, etc. This thesis presents the very first trial to correlate EGFR movement within a cell using these techniques