Image-Based Graph Analysis to Capture F-actin Cytoskeleton Morphological Changes

The morphological and mechanical properties of the cytoskeleton are critical for understanding structural changes in cells, whether driven by disease progression or
external interventions. External factors influencing the cytoskeleton include pharmacological agents such as cytochalasin, while pathological conditions include
cancer, and physical stimuli like photobiomodulation. Numerous studies have explored mechanical alterations, but quantitative analyses of cytoskeletal
morphology remain limited. In this study, we quantitatively investigate the morphology of the F-actin cytoskeleton in multiple cell lines exposed to various
external perturbations, employing a graph-based analytical approach. Confocal fluorescence microscopy images of actin filaments are used to construct graph
representations of the cytoskeletal network, enabling detailed structural characterization. We begin by examining correlations between graph-derived
features—such as network topology, connectivity, and filament organization—and biological aspects of the cytoskeleton. Subsequently, we analyze how these graph
metrics vary under different perturbation conditions, including cytochalasin exposure time, red light irradiation, and comparisons between cancerous and healthy cell
lines. Finally, we relate the observed morphological changes to previously reported mechanical alterations, offering a more integrated understanding of cytoskeleton
remodeling under diverse influences. This quantitative framework provides new insights into the structural dynamics of the cytoskeleton and advances our
understanding of cellular responses to both biochemical and physical stimuli.