Engineering Vitrification Methods for Nanoscale Visualization of Dynamic Cell Processes

Abstract

Biological visualization is a critical tool for understanding and communicating biological processes, mechanisms, and interactions. A key gap in current visualization technology is the simultaneous imaging of processes that take place in nanoscale spatial regimes and millisecond temporal regimes. This work addresses these gaps through the development of a new tool for preparing biological samples, in addition to improvements in the sample preparation itself to allow access to a range of previously inaccessible thicker samples. Additionally, we address the development of biological cell lines expressing proteins crucial to human health and physiology, and the subsequent imaging of fluorescently tagged protein constructs we express in these cells. The data analysis to process these images is nontrivial, and we therefore develop new algorithmic techniques for analyzing the colocalization of two protein species in high density and noisy experimental contexts.