Re: nuclear stain

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From:Geoff McAuliffe <mcauliff@UMDNJ.EDU>
To:Heike Grabsch <>
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Heike Grabsch wrote:

> Hi Histonetters,
> I am sure there is somebody out there who is able to help us! WE do
> double immunhistochemistry labeling, one chromogen is DAB+copper sulfate
> (nively black) for detection of a nuclear antigen, the other is AEC-one
> step solution from Biogenex which gives you a real wonderful red fro a
> cytoplasmic antigen. However, I would like to counterstain all other
> nuclei. I thought about methyl green, however either it is alcohol-based
> like in Romeis (germany histotechnology book) or need to be at least
> dehydrated via n-butanol and xylol linke in John Kiernan's book. But
> alcohol will kill my AEC-staining! Any other green nuclear stain or may
> be yellow which is non-alcohol-based and does not need to be dehydrated?

How about dehydrating with acetone or dioxane or tetrahydrofuran or 2.2
dimethoxypropane? Perhaps AEC will survive one of those reagents. Or just
air dry and then go to a clearing agent?

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

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