Posted by
Michael Giacomelli on
URL: http://confocal-microscopy-list.275.s1.nabble.com/fixable-dead-cell-nuclear-counterstain-tp7586389p7586392.html
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All dyes I have used are greatly attenuated by xylene processing,
however some remain reasonably bright. Propidium iodine is a good
choice, although unless you deliver a reasonable amount it may be
quite dim post processing.
Mike
On Wed, Feb 1, 2017 at 9:25 AM, Dave Johnston <
[hidden email]> wrote:
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>
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>
> Can anyone recommend a non membrane permiable, fixable, nuclear counterstain, which neither fluoresces far red, nor requires hard UV excitation (so DAPI is out) which can then withstand wax embedding for histology? We have a user who wants to test a novel delivery system into which an aqueous phase dye can be incorporated, the test being that if the delivery system gets into a cell and is processed, the dye will be released and enter the nucleus to bind the DNA and stain the cell. The experiments will be in vivo, hence there is a need to fix and wax embed.
>
> I know that there are membrane impermeant, fixable cytosolic amine reactive dyes which would stain the cell if delivered this way but wondered about a nuclear stain.
>
> There seems relatively little literature on fixable DNA dyes or does one assume that aldehyde based fixation will cross-link any already incorporated dye.
>
> Thanks in advance,
>
> Dave Johnston,
> Biomedical Imaging Unit,
> Southampton.