[Histonet] whole mounts

From:"Till, Renee"

Does anyone have experience with whole mount slides, specifically
mammary whole mounts? I have done a couple batches of rat mammary whole
mounts with varying results. The protocol I follow is pretty simple.
They are fixed, then defatted in acetone, rehydrated in 70% ETOH and
water, stained in carmine, then dehydrated through several alcohol
gradients, and cleared in xylene. Before coverslipping I also flatten
the tissues by pressing them between two slides.  Depending on the
quality of tissue collection, most of my slides turn out well. There are
usually a couple that the stain is dark and almost black instead of
purple and I cannot use these. 
I guess my questions are, beyond just any general advice, does anyone
have a different (perhaps better?) protocol and do you think the bad
ones could be destained and redone? I'm of the opinion that it probably
can't be done, but I keep getting asked by my boss. 
I'm pretty much pleased with the results myself, but if it can be done
better...It seems my biggest problem is publication quality pictures to
go with the data I generate off these slides. Our equipment is good, and
in the microscope I can often see the structures, but some stain more
darkly than others and the lighter ones just don't show up. Of course
these are the ones I need good photos of. 
Any suggestions?  Is there a better stain? 
Renee' Till
Arkansas Childrens  Nutrition Center 


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