fixable, dead cell nuclear counterstain.

Posted by daj1u06 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/fixable-dead-cell-nuclear-counterstain-tp7586389.html

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

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.