[Histonet] Purple haze on H&E slides

[Blazek, Linda]

Haley,
 We use Leica's Selectech also and occasionally have the same issue.  I don't think your ETOH after the eosin is a problem.  If you add a 95% you will leach out the eosin a bit.  I think you should add another 15 - 30 seconds in the acid alcohol.  One of the things we find here though is that there can be a great variation to the pH of the water.  That affects the results of the stain. Specifically the "purple haze"  Go Jimi!

Purple haze all in my brain 
Lately things just don't seem the same 
Actin' funny, but I don't know why 
'Scuse me while I kiss the sky 

Linda
-----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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