[log in to unmask] wrote:

>The improvement in resolution, even with thin specimens, is caused by
>selective illumination at the points of interest, and happens even without a
>confocal aperture.  Image from one volume within the specimen is not degraded
>by stray light from other regions, since the rest of the specimen is dark
>most of the time.

Thanks, that makes sense indeed.

Beat wrote:
>> + illumination confined the the scanning plane, so there should be less
>> phototoxicity problems.

David replied:
>Note however that while illumination is focussed at one plane, it passes
>through all others, so photobleaching can occur in planes which are not under
>observation...

Steve replied:
>Is this really true?  How is the illumination confined to the scanning plane?

Sorry, of course my statement as such was just plainly wrong (I should
re-read my late nite writings before I send them off in the morning ;-) I
have been discussing too much about combining the CSU10 with a 2-photon
system, I guess.
However, because the relation between illumination density/duration and
bleaching is not always proportional, selective bleaching in the focal
plain can occur (and I had the impression that the bleaching in my
samples was largely confined to the focal plane in my tubulin-GFP
samples).

Thanks for your comments,

Beat



-----------------------------------------------------
Dr. Beat Ludin, FMI, Maulbeerstr 66, 4058 Basel, Switzerland
Tel. +41 61 697 6697 / FAX +41 61 697 3976
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