Photobleaching during live cell imaging experiment


I am interested in doing some SIM live-cell imaging on OMX. One of the obvious problems that I am going to encounter is photobleaching of my sample. Apart from things that I can control (laser power, exposure and interval between images) to minimize photobleaching, has any one tried using prolong live antifade from ThermoFisher.


Sorry about being so late on this. How long do you need to image? If it is short times (less than 1 hour, or a single acquisition for example), then using a live cell incubation buffer might help, here, you may be able to significantly drop the laser intensity while achieving the same signal to noise, and thereby minimize photobleaching. I’m not sure what the Thermo product is, but people have been adding anti-oxidants/radical scavengers to imaging media for some time, it is purported to make a difference in photobleaching. Good luck with your experiments.