Microtubule dynamics with optimal image settings. LLC-PK1 cells stably expressing EB3-mEmerald and H2B-mCherry were seeded onto fibronectin-coated dishes. Cells were simultaneously illuminated for ~2 mins with a 491 nm laser set to 0.020 mW to image mEmerald and a 561 nm laser set to 0.010 mW to image mCherry. Images were captured with an HCX PL APO 63x/1.40NA oil objective and camera exposure time of 200 ms. mEmerald and mCherry emission signals were sent to two separate sCMOS cameras using a dichroic mirror, then colour coded and digitally combined. Time is in min:s. Scale bar is 5 μm.