High‐Content Analysis of Cytoskeleton Functions by Fluorescent Speckle Microscopy

Abstract The development of nanomedical strategies requires precise mechanistic models of cellular pathways. At center stage of many of these pathways is the cytoskeleton, which dynamically organizes cellular architecture and mediates cell morphological functions. In this chapter we review the principles and selected applications of quantitative fluorescent speckle microscopy (qFSM), a combined imaging modality and statistical toolset that delivers high‐resolution measurements of cytoskeleton structure and dynamics in living cells. qFSM has emerged as an invaluable tool in studies aimed at delineating the mechanisms of cytoskeleton regulation by the systematic perturbation of molecular signals upstream of cytoskeleton outputs. Additionally, qFSM can be used as a precise readout of cytoskeleton responses in the search for new drugs and nanomedical strategies that alter cellular behavior by intervening with specific cytoskeleton functions.