[Histonet] H&E Stain

koellingr <@t> comcast.net koellingr <@t> comcast.net
Thu Jan 20 13:07:29 CST 2011

Thought I'd throw this out. Maybe nonsense. If you have such acidic tap water, could there be heavy metals lead, magnesium and others (acid tap water does that) in your tap water rinse from being leached out upstream. William DeSalvo talked about the quality of tap water fluctuating. Very true. And the metals from pipes or solder , leached into water by pH6.0, turning a normal hematoxylin into something like a Weigerts hematoxylin. A kind of post-mordanting that I think some call afterchroming. Although if you tried distilled or deionized water with same results, that data wouldn't fit with this problem. And even if it just started happening, has someone recently worked on pipes upsteam of where you are and there is (are) new metals being leached into your hematoxylin rinse? pH 6 is pretty acidic water. 

PhenoPath Labs 
Seattle, WA 

----- Original Message ----- 
From: "Toni Rathborne" <trathborne <@t> somerset-healthcare.com> 
To: "WILLIAM DESALVO" <wdesalvo.cac <@t> hotmail.com>, jkiernan <@t> uwo.ca, "allison scott" <allison_scott <@t> hchd.tmc.edu> 
Cc: "histonet" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Thursday, January 20, 2011 6:17:46 AM 
Subject: RE: [Histonet] H&E Stain 

Our tap water consistently reads 6.0, and has for years. We did try turning off the tap when this first began, and manually rinsing with distilled water, but saw no difference. I will try adding the HCl today with a few test slides. Will let you know how this works out. Thanks for the suggestions. 

-----Original Message----- 
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of WILLIAM 
Sent: Thursday, January 20, 2011 12:26 AM 
To: jkiernan <@t> uwo.ca; allison_scott <@t> hchd.tmc.edu 
Cc: histonet 
Subject: RE: [Histonet] H&E Stain 

I agree that the pH might be high, but I also suggest you check your water rinse on the stainer. If you are using "tap" water, there can be a significant fluctuation in the quality of the water and the amount of additives and impurities present at any one time can also contribute to the mucin not being rinsed away and staining. If you are using tap water, changing to distilled or dionized water might help to improve the consistency of stain results. Good luck 

William DeSalvo, B.S., HTL(ASCP) 

> From: jkiernan <@t> uwo.ca 
> To: Allison_Scott <@t> hchd.tmc.edu 
> Date: Wed, 19 Jan 2011 16:58:57 -0500 
> Subject: Re: [Histonet] H&E Stain 
> CC: histonet <@t> lists.utsouthwestern.edu 
> Sounds as if the pH of your haemalum is too high. Try adding a little HCl to bring it down to slightly above 2. Check a few slides without eosin counterstaining. Nuclei should be blue with very little else stained. 
> John Kiernan 
> Anatomy & Cell Biology 
> University of Western Ontario 
> London, Canada 
> = = = 
> ----- Original Message ----- 
> From: "Scott, Allison D" <Allison_Scott <@t> hchd.tmc.edu> 
> Date: Wednesday, January 19, 2011 13:01 
> Subject: [Histonet] H&E Stain 
> To: histonet <@t> lists.utsouthwestern.edu 
> > Hello to all in histoland and Happy New Year. We are 
> > having issues with 
> > our H&E stain. The nuclei are staining very blue to purple 
> > and the 
> > mucin is staining blue to purple-blue. It is difficult to 
> > see the 
> > nuclear detail. The mucin is obscuring things. We 
> > have not changed our 
> > process for staining or processing. The funny thing is 
> > that it is only 
> > in the Biopsy cases, and it is every few slides. The 
> > surgical cases 
> > are all right. We checked the alcohol and xylene for 
> > water, and there 
> > is not any. My tech changed out the stain and we are 
> > staining a new 
> > batch of slides. If anyone has any idea what is wrong, any 
> > help would 
> > be greatly appreciated. I have gone over our processes and 
> > nothing has 
> > changed. The reagents are the same, the staining times are 
> > the same, 
> > and the processing times are the same. We are using the 
> > Shandon Gemini 
> > stainer and VIP processor. 
> > 
> > Allison Scott HT(ASCP) 
> > Histology Supervisor 
> > LBJ Hospital 
> > Houston, Texas 
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