Label-free cargo localization iSCAT microscopy uncovers dynamic transport of intracellular vesicles and underlying active cytoskeletal network

We introduce the cargo-localization iSCAT microscopy, label-free, live-cell imaging technique, that visualizes the traffic of cargos in a massively parallel fashion and hence uncovers the actively-used cytoskeletal network from cargos’ traces. By applying a particle localization algorithm to over a hundred cargos moving in a lamellipodium, the fine architecture of the cytoskeletal highway could be reconstructed. We also discuss a host of interesting kinetic events associated with cargo transport such as intermittent pausing, turnover, and frequent jamming of cargos due to physical collision with other obstacles or at the intersection of the highway.