Re: Photobleach (FRAP) problem in Zeiss 510Meta

Posted by Chen, Mei Ling on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photobleach-FRAP-problem-in-Zeiss-510Meta-tp5038048p5046766.html

Dear Guosheng,
Did you make the right registration of the ROI on Bleach edit and Time
series control windows?
We have had very good and stable results of FRAP from LSM510 and LSM510
META microscopes.
Mei Ling

On Tue, May 11, 2010 3:21 pm, G.Liu wrote:

> 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
>
>
>
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