Dear All,

Thanks you very much for all of your input (comments, suggestions, protocols
etc ) in regard to the photobleaching in our Zeiss510. The good news is that
we got some progress on this but bad news is that a new question needs to be
addressed: the movement of the bleaching subject/organelle. 

We are using transiently GFP-expressed tobacco leaves and want to bleach
whole or partial nucleus. However the nucleus seems to move at lease about
1-2um in a 5min period. we tried to use larger pinhole and larger ROI but
the plane of focus is still an issue (so only saw partial bleach). Zeiss
specialist suggest to stick the leaves on the slide with krazy glue and
surrounded by something to avoid the leave floating/vibrating (we are using
inverted microscope). We tried but the movement was still noticeable. Have
some of you meet a similar problem and how did it solved? 

Meantime I am want to install a macro that could track the cell movement
(developed by Ellenberg). Any experience on it?

Thank you in advance.
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