Fwd: [Histonet] H&E Staining

[godsgalnow <@t> aol.com]

-----Original Message-----
From: godsgalnow <@t> aol.com
To: Kemlo.Rogerson <@t> waht.swest.nhs.uk
Sent: Wed, 10 Oct 2007 11:10 am
Subject: Re: [Histonet] H&E Staining


How long are you in the post-eosin staining?? I would skip the 70 % and go straight to 1 change of 95% and 3 changes of 100%.

Roxanne


-----Original Message-----
From: Kemlo Rogerson <Kemlo.Rogerson <@t> waht.swest.nhs.uk>
To: William Ares <wia2005 <@t> med.cornell.edu>; histonet <@t> lists.utsouthwestern.edu
Sent: Wed, 10 Oct 2007 9:40 am
Subject: RE: [Histonet] H&E Staining



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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