Visualizing Filamentous Actin in Chlamydomonas reinhardtii

: This protocol aims to visualize the filamentous actin network in Chlamydomonas reinhardtii. We improved fixed-cell labeling conditions using the F-actin probe, phalloidin. We created a Chlamydomonas-optimized protocol by halving the phalloidin incubation time, electing for optimal fixation conditions, and selecting for a healthy cell population. This phalloidin protocol is quick, effective, and is the only labeling method to date that allows for reliable actin filament detection in fixed vegetative Chlamydomonas cells. This method reveals previously unidentified actin structures in Chlamydomonas and novel insights into cytoskeletal dynamics.