Hello all,
I perform immunohistochemistry (IHC) and immunofluorescence (IF) on
fixed free-floating rat brain sections (~50um thickness). Currently I
use Prolong Gold with DAPI as a mounting media as well as a stain for
the nuclei.
I am seeking an alternative (fluorescent) nuclei stain for free floating
rat brain sections. The issue I am having is that one of my fluorescent
markers (FluoroGold) has native fluorescence in the UV channel. Since
it is the same channel that DAPI is in, this will not do. I do not have
an antibody or fluorescent marker in the far-red channel so I would
really like to find one in that area.
I have looked at To-Pro-3 and NucRed Dead 647 as alternatives because
their fluorescence is in the far-red channel. However I am unclear if
these stains can be used on brain sections and not just in vitro cells.
To summarize, I am looking to find an alternative nuclei stain to DAPI,
and one that is preferably in the far-red channel. If anyone here has a
protocol for a nuclei stain that they would not mind sharing, I would be
very appreciative as well.
Thank you so much for your help,
Mike Bruno
State University of New York at Buffalo
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