Imaging Mammalian and Dictyostelium Cells

All mammalian cells were cultured in DMEM (ATCC, #30-2002) supplemented with 10% fetal bovine serum (ATCC, #30-2020) at 37 °C and 5% CO₂, and all imaging of mammalian cells was conducted under serum free conditions (DMEM alone). Hs27 fibroblasts (ATCC, #CRL 1634) were imaged on planar sections of quartz contact guidance chips as previously described^{28 (link)}. MDA-MB-231 cells (ATCC #HTB-26) were imaged on glass bottomed well plates coated with 25 µg/mL Fibronectin (Gibco #33016015) or functionalized gold coated coverslips as previously described^{29 (link)}. A549 cells (ATCC #CCL-185) were imaged on planar sections of quartz nanostructured chips as previously described³⁰ . The Dictyostelium cells were wild type AX2 strain and generously gifted from the Devreotes laboratory of Johns Hopkins University, were cultured axenically in HL5 at 22 °C, and imaged on glass bottom well plates as previously described^{31 (link)}. Microscopy details for each cell type including microscopy mode, magnification, numerical aperture, camera and wait time between images are listed in Supplementary Note 2.