[Histonet] RE: Eosin not working for xylene free H&E

[Goins, Tresa]

We use hot detergent solution for dewax and a xylene substitute (Pro-Par) after staining without problems.  If there is water accumulating in the clearant, it can be removed with a molecular sieve placed in the staining trays.  We don't have that problem here in Montana because it is dry dry dry.

Tresa 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sarah Dysart
Sent: Wednesday, March 27, 2013 8:11 AM
To: Gerard Spoelstra; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Eosin not working for xylene free H&E

This exact reason is how I convinced my lab to go back to xylene!!  I figure it's because there is water in your substitute.  Because you can't see if there is water in it (from the air moisture or wherever...) it becomes frusterating and annoying to have to change out to fresh solutions sometimes several times a day!!  
Good Luck!!  


Sarah Goebel-Dysart, BA, HT(ASCP), QIHC (ASCP) Histotechnologist Mirna Therapeutics
2150 Woodward Street
Suite 100
Austin, Texas  78744
(512)901-0900 ext. 6912

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gerard Spoelstra
Sent: Wednesday, March 27, 2013 9:07 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Eosin not working for xylene free H&E

Hi everyone,

I have not had any success optimising the eosin counterstain in the absence of xylene. We routinely use 0.5% eosin Y in 95% ethanol. I've tried taking sections through to this to absolute ethanol but then air drying , but it leaves blotches of eosin.  Using eosin 0.5% aqueous with a few drops of acetic acid doesn't appear to work, the keratin stains heavily but not elastin. After washing in water I leave for a short time in the oven then at room temperature for 5 minutes then coverslip with the dako atomatic cover slipper.   For all the laboratories that have gone xylene free, are you just putting up with less than optimal eosin staining?

Gerard Spoelstra
Murdoch University
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