Put a few drops of glacial acetic acid in the eosin. Without 
acidification, the stain will not 'adhere' to tissue components.

Geoff

Kemlo Rogerson wrote:
> i'm currently trying some H&E staining (post-TUNEL) on paraffin embedded
> tissues and keep having the eosin wash out during the dehydration steps
> (stepwise: 70% alcohol, 95% alcohol, 100% alcohol)... any ideas on why
> this would happen?  am i missing a step in between that allows the stain
> to 'stick'?  thanks
>
> It is 'stuck' but you are unsticking it. Either because you are using
> eosin that is soluble in alcohol or because the water in the 70% is
> removing the water soluble eosin. You could try adding calcium ions as
> an accentuator, that might make the glue stronger, but of course you
> must use aqueous eosin or the calcium ions come out of solution.
>
> Kemlo Rogerson
> Pathology Manager
> DD   01934 647057 or extension 3311
> Mob 07749 754194; Pager 07659 597107;
>  
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-- 
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583 
mcauliff@umdnj.edu
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