Figure 12. Using 2 sequential acquisitions 15 seconds apart to ensure that the acquisition has not modified the membranes or damaged the cells.

Hela cells grown for two days on glass coverslips were stained with 200 nM C-laurdan and imaged twice sequentially with the standard conditions defined in M&M (power 4%, speed 6, 4 averages, pinhole 150). With these setting, the acquisition procedure takes 15 seconds. The second picture is thus taken 15 seconds after the first one. Z4:Whole image : 53 µm, 1 pixel = 100 nm HELA C Laurdan 200 nM, Laser at 720 nm, Z4, power: 4%, speed: 6 (6 usec/pixel), average on 4 measures, pinhole: 150 Hela CL 4 and Hela CL 5 15 Oct 2015.