Measure particle diffusion in cytoskeletal networks with light-sheet and differential dynamic microscopy

Filamentous actin and microtubules contribute to the crowded environment of the cytoplasm. Modeling the cytoplasm, we create networks of varying crosslinking motifs and varying ratios of actin and microtubules. We crosslink either actin to actin, tubulin to tubulin, or tubulin to actin. We add micron-sized fluorescent beads to the various networks, record videos of the samples using light-sheet microscopy, and perform differential dynamic microscopy (DDM) analysis. We compare DDM and single-particle tracking results to examine differences between ensemble and single-particle transport properties. We find varying degrees of anomalous diffusion and significant heterogeneity in the dynamics, particularly in actin environments.