[Histonet] H&E Staining

[Jennifer MacDonald]

Also, the more water that  is in the dehydrating alcohols, the faster the 
slides will differentiate.  You may be washing out the eosin.  The optimal 
pH is 4.6 to 5.



Jennifer MacDonald
Director, Histotechnician Training Program
Mt. San Antonio College
1100 N. Grand Ave.
Walnut, CA 91789
(909) 594-5611 ext. 4884
jmacdonald <@t> mtsac.edu



Geoff McAuliffe <mcauliff <@t> umdnj.edu> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
10/10/2007 08:23 AM

To
Kemlo Rogerson <Kemlo.Rogerson <@t> waht.swest.nhs.uk>
cc
William Ares <wia2005 <@t> med.cornell.edu>, histonet <@t> lists.utsouthwestern.edu
Subject
Re: [Histonet] H&E Staining






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
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