I wanted to observe the chloroplast's response in a thallus tissue after photobleaching in FRAP. But, the observation focus always shift away and eventually the observation is lost while the timelapse were taking. May I have any suggestions to avoid this problem?
My setting in the FRAP
> Argon- 20%
> WL- 70%
> Scan mode- xyt
> Sequential scan- 1
> Pinhole size- 1
> Bidirectional scan- on
> Zoom in (2)- on
> Set background to zero- on
> Use laser settings for all ROIs - on
> Repetitions- 100 (ratio for pre-bleach to bleach to post-bleach- 1:2:1)