Movie 1

Cells imaged with short exposure times migrate significantly slower than cells imaged with longer exposure times when the light source is activated through USB. CHOK1 cells stably expressing paxillin-EGFP were seeded onto fibronectin-coated plates and allowed to migrate for 3 hours. Images were captured with a Plan ApoChromat 20x/0.8NA objective. For continuous fluorescence illumination (DLD), a 120LED light source was used to deliver 7% intensity through a 10% ND filter (OD=1) and a 1% ND filter (OD=2) for a final intensity of 0.007% (0.0245 mW); camera exposure time was set to 60,000 ms. For CLD, 21% incident light source power (61.6 mW) was delivered to the sample; camera exposure time was set to 24 ms. Microscope software was used to activate the light source through a USB connection. Time is in hr:min. Scale bar is 25 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