Re: Photobleach (FRAP) problem in Zeiss 510Meta

Posted by Axel Kurt Preuss on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Photobleach-FRAP-problem-in-Zeiss-510Meta-tp5038048p5039119.html

It could be it s not aligned and you are bleaching in a different spot
Best to take a fluorescent slide and test xy

Thanks

Axel

Axel K Preuss PhD, A*Star IMCB-Central Imaging Facility 6-19B   Sent
from. [phone]

On May 12, 2010, at 4:19 AM, "G.Liu" <[hidden email]> 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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