Posted by
Glen MacDonald-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Background-fluorescence-problem-tp7579031p7579033.html
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Dear Simon,
there are reports of autofluorescence arising from flavins and aromatic amino acids in culture media additives. This apparently varies among different lots of additives. these compounds have been also implicated in light-dependent toxicity. Ch. 19 of The Handbook mentions this, with some references.
If the fluorescence recovers, I'd suspect that it is replenished by media circulation.
Regards,
Glen
Glen MacDonald
Cellular Morphology Core
Center for Human Development and Disability
Box 357920
University of Washington
Seattle, WA 98195-7920 USA
(206) 616-4156
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On Sep 13, 2012, at 8:04 AM, Simon Walker wrote:
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> Dear List,
> We are imaging very weakly fluorescent live cells (expressing GFP) on a wide-
> field system and having issues with a source of background fluorescence.
> When we look at our cells under epi-illumination we see a rapid drop in a weak
> background signal (not where the cells are) that fully recovers over a ~10 s
> period after the illumination light is switched off. Our experiments require the
> use of DMEM as the imaging medium and this is the likely cause of problem. It
> appears that something in the medium is sticking to the coverglass. It's not
> phenol red as the effect is seen with both phenol red-containing and phenol-
> red-free DMEM. Does anyone know what else it could be? Has anyone else
> seen anything similar? We're wondering if it could be riboflavin which is in the
> DMEM we're using. Would this stick to glass?
>
> I've seen that Life Technologies now market a substance that allegedly
> surpresses background fluorescence in DMEM:
>
http://products.invitrogen.com/ivgn/product/R37603> Has anyone tried this? Does anyone know how it works?
>
> Thanks,
> Simon