21 December 2021 0 6K Report

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)

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