Movie 6

Continuous imaging reveals highly dynamic adhesions. Lung-metastatic 4T1 breast cancer cells (4T1-537) were seeded onto fibronectin-coated dishes and transfected with mCherry-paxillin. Cells were continuously illuminated for 20 mins with a 561 nm diode laser set to 0.031 mW. 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 the protruding edge of the cell (white box). Every sixth image was copied into a new image stack to illustrate the effect of only collecting one image every 30 s (red box). Time is in min:s. Scale bar is 5 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