Posted by
George McNamara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/DAPI-weirdness-with-hardset-Vectasheild-tp6869170p6873024.html
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Yes, I have seen DAPI, as well as Hoechst (33342, 44258) photoconvert to
green. This has been previously discussed on the listserv.
To avoid this, try using a much lower concentration, incubate the
counterstain, then wash, then apply mounting medium. Avoid using old
mounting medium. Change to Prolong Gold (without DAPI).
On 10/7/2011 6:51 AM, Simon Walker wrote:
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> Has anyone else observed weird behaviour of DAPI when used in conjunction
> with hard-setting Vectasheild? When exposed to wide-band UV excitation we
> have seen the DAPI excitation characteristics change such that the DAPI
> becomes excitable with 488 nm light. And today someone showed me a slide
> where (again using wide-band UV excitation) the DAPI signal initially looked a
> pinky/red colour, but after a few seconds of exposure developed the usual
> blue fluorescence. We don't seem to have these problems with the liquid
> mount Vectasheild. Any suggestions as to what might be going on??
> Thanks,
> Simon
>
>
--
George McNamara, PhD
Analytical Imaging Core Facility
University of Miami