Photobleach (FRAP) problem in Zeiss 510Meta

Posted by G.Liu on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photobleach-FRAP-problem-in-Zeiss-510Meta-tp5038048.html

Dear all,

We have encountered a problem in our 510Meta for photobleaching. We simply could not get the GFP ROI bleached. What we did is as following, anything wrong or inappropriate?


1. Collect the Single unsaturated image as usual (normal settings).

2. Go to Bleach control for bleach settings. Select xx scans before the bleach. For GFP, tried 1-50 iteration, defined the Bleach region. All the argon laser lines (458, 488, 514) or only 488 to 100% .

3. Go to Time Lapse--using Manual start, manual stop (input the number of scan). Time interval 0 or 1 sec.

4. Hit StartB.

I can get a series of images but no bleaching was found. The other odd thing is that "Bleach" button seems not not working--if hit, it only lasts a few seconds then stop. I've tried using diode405 line, it worked once, but I can't repeat it anymore.

Any suggestions or advice are greatly appreciated!!

Guosheng