Modified protocol for culturing Drosophila S2 R+ cells and adult plasmatocytes to study actin cytoskeleton dynamics

Here, we describe a protocol to culture Drosophila S2R+ cells and to extract plasmatocytes from adult flies. The modified seeding approach detailed here, in combination with coating of coverslips with concanvalin A, enables enhanced adhesion and spreading of cells. We describe the steps for confocal microscopy and a detailed quantification pipeline to evaluate changes in cortical actin cytoskeleton dynamics. The protocol can be applied to a variety of genetic or chemical perturbations. For complete details on the use and execution of this protocol, please refer to Nath et al. (2022).