Movie 7

Continuous imaging provides superior temporal resolution for imaging actin cytoskeleton dynamics. NMuMG cells were infected with LifeAct-EGFP and seeded onto fibronectin-coated dishes. Cells were continuously illuminated for 30 mins with a 491 nm diode laser set to 0.060 mW to image EGFP fluorescence. Images were captured with an HCX PL APO 63x/1.40NA oil objective and camera exposure time of 5 s. 2x2 pixel binning was used to enhance S/N. Time series was cropped to zoom in on part of the protruding edge (white box). Time is in min:s. Scale bar on inset video series is 3 μm.

Optimizing live-cell fluorescence imaging conditions to minimize phototoxicity

Alex Kiepas, Elena Voorand, Firas Mubaid, Peter M. Siegel, and Claire M. Brown

J Cell Sci 2020. 133:None-None; doi: 10.1242/jcs.242834