[Histonet] Suggestions for alternative fluorescent nuclei stains (that are not DAPI) ?

[Michael Bruno]

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