Topological Analysis of Cytoskeletal Networks from Single-Molecule Localization Microscopy Images

Abstract

The cytoskeletons are fundamental structural elements of the cells, permeating the cell interior and playing essential roles in many cellular activities. The architecture of the cytoskeletons is a critical physical attribute governing their biological functions. However, challenges remain in the quantitative understanding of the cytoskeletons. In this dissertation, I addressed these by an integrated approach combining super-resolution microscopy with advanced computational analysis. First, a systematic optimization of single-molecule localization super-resolution microscopy was performed. This enabled the visualization of the cytoskeletons at nanoscale resolution. Second, novel computational approaches were developed for the quantitative analysis of the intricate cytoskeletal networks from super-resolution images. This enabled automated high-throughput analysis of the cytoskeletal filaments from complex images. These advances were demonstrated in the studies of microtubule network organization downstream of the small GTPase Rac1, and the Sertoli cell cortical F-actin network organization downstream of the actin polymerization factors, mDia1 and mDia3.