[Histonet] Purple haze on H&E slides

[Anita Buchiane]

It looks to me like the problem might be caused by inadequate dehydration after the eosin.  Add a 95 % after the eosin then 3 100 %'s then the Xylenes.  2 min. in all of these should be adequate.
Hope this helps

Anita
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Huggins, Haley - MRMC
Sent: Tuesday, February 11, 2014 12:07 PM
To: Campbell, Tasha M.
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Purple haze on H&E slides

We are using the Leica Selectech brand. No additives to the waterbath. We are using xtra or superfrost slides. The protocol is as follows:
3 min xylene
3 min xylene
1 min 100% ETOH
2 min 100% ETOH
1 min 95% ETOH
1 min water rinse
4 min hematoxylin
2 min water rinse
15 sec acid alcohol
1 min water rinse
1.5 min bluing
1 min water rinse
1 min 95% ETOH
5 min eosin
30 sec 100% ETOH
1 min 100% ETOH
1 min 100% ETOH
1 min xylene
2 min xylene

Haley

-----Original Message-----
From: Campbell, Tasha M. [mailto:tmcampbell <@t> fmh.org] 
Sent: Monday, February 10, 2014 5:38 PM
To: Huggins, Haley - MRMC
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Purple haze on H&E slides

What brand of reagents are u using and what is your protocol? What slides r u using and r u using gelatin in ur waterbath?

Tasha

Sent from my iPhone

> On Feb 10, 2014, at 7:10 PM, "Huggins, Haley - MRMC" <Haley.Huggins <@t> DignityHealth.org> wrote:
> 
> I am trying to get the histology lab up and running and I am having difficulties getting the H&E Stain just right. I am nearly there, but I keep getting a purple bleed or haze around the tissue. Any suggestions on getting rid of this? I actually got rid of it at one point, but the medical director wasn't happy with the stain. I appreciate any suggestions from the histology world.
> 
> Haley Huggins
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> Histonet <@t> lists.utsouthwestern.edu
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