[Fluorescent-proteins] bars across field of view
(by heterochromatic from gmail.com)
Thu Aug 19 15:51:18 EST 2010
I wonder if anyone out there can explain an image problem I am having
with my fluorescence microscopy. I am currently using a Zeiss Axioscop
but I have seen the same thing on a Nikon some years back, so think
this might be independent of brand.
1. Dark bars, like a fuzzy grid, appear across the field of view.
2. They are only visible when there is broad fluorescence throughout
the field of view. Ie., I do not see them in the DAPI channel b/c the
image is mostly black space with lots of bright blue spots. But I see
it in the GFP channel if I am looking at ALOT of fluorescence
3. So far, I have only seen this on the GFP channel, but then that is
the only channel I have that showed that broad fluorescence (the scope
is new - the red and blue have been more defined localizations based
on the antibodies/dyes). But I suspect that if I had lots of rhodamine
staining, I would see it on the red channel also (it was with
rhodamine I remember seeing it on the Nikon).
4. I see it through the 40X and 60X lenses, but not the 10X (and I
haven't got a 100X yet).
5. It does not move with the slide, so it isn't on the cover slip. In
any case, it appears over multiple samples.
6. I have examined the filter cubes and they are spotless.
7. I have cleaned all the lenses, thinking there may be a fingerprint.
But the bars are still there.
8. The light source for this scope is LED (Zeiss's colibri system, not
an arc lamp) and as per the advice of the tech I have rotated the
modules, no change in the orientation of the bars. (Anyway the Nikon
was not LED, it was Hg.)
I would like to attach an image but I think that might not be allowed,
so if someone knows what I am talking about, please could you contact
me and I will send an image. I would like to get this sorted out while
the scope is still under warranty (couple of months left...)
Western Washington University
Bellingham WA 98225
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