[Histonet] Routine H & E stain,

[Rittman, Barry R]

I believe that this could be a multifactorial problem.
There is a tendency for us, "in the interests of efficiency" to process tissue and slides as rapidly as possible. In a lot of cases I suspect that the absolute minimum time in reagents causes some carry over that can result in fading of the stains.
The quality of the xylene, as one person already pointed out may be a factor. Even using "real xylene", the composition, sulfur content etc can vary considerably and affect staining. There was some concern in the 1950s over fading of slides containing silver stains due to using xylene. However I still have some stained with silver that were cleared with xylene and still look really good.
The use of xylene substitutes has not (to my admittedly imperfect knowledge) been tested over a few decades.

It would also be interesting to know whether the type of hematoxylin stain used has any effect on fading of hematoxylin. I have always been an advocate for Ehrlich's hematoxylin and have 50+ year old slides that  are still as bright as the day they were stained but not much experience with other hematoxylins.

Mounting media may also have an effect. In the days of the dinosaur we used Canada balsam in xylene and later, DPX. Slides mounted in these still look really good, in fact dare I say beautiful.

Might be an idea to use some pH paper to check pH of the xylene and mountants.

Some have also suggested that exposure to UV light may also be a factor. Not sure that there is much evidence for this.

I have seen some slides from the mid 1800s that while not pristine still have some hematoxylin staining. 

Bottom line, is that we have no bloody idea what is causing your hematoxylin to fade- suggest that failing all else, you pray and enjoy a glass of Zinfandel (Paso Robles 2006 was a really good year).
Barry

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Thursday, July 07, 2011 9:45 AM
To: Histonet; histonet-bounces <@t> lists.utsouthwestern.edu; histonet; amitapandey <@t> torrentpharma.com
Subject: Re: [Histonet] Routine H & E stain,

It seems that your mounting medium is acid. Try to correct that rather than changing the staining.
René J.

--- On Thu, 7/7/11, amitapandey <@t> torrentpharma.com <amitapandey <@t> torrentpharma.com> wrote:


From: amitapandey <@t> torrentpharma.com <amitapandey <@t> torrentpharma.com>
Subject: [Histonet] Routine H & E stain,
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>, histonet-bounces <@t> lists.utsouthwestern.edu, "histonet" <histonet-request <@t> lists.utsouthwestern.edu>
Date: Thursday, July 7, 2011, 12:22 AM


Hello Histonetters,

I am from toxicopathology lab where we perform H&E on rat tissues and 
store these slides for 10 long years.

We use self lab prepare Harris hematoxylin (Hematoxylin crystal+Alcohol+ 
Ammonium alum +D/W and Mercuric oxide for ripening) and 1% aqueous eosin. 
The staining result is good (Purplish pink look stained slide) on all 
tissue, but  our observation is after 5-6 months time this get faded and 
become towards pinkish type though we can observe the slide. 
We want to retain its fresh stained color. 

Please suggest me how to keep stable stain for longer period or do you 
suggest to switch to any other type of haematoxylin - prepared or 
commercially available?

Looking forward for your feed back.
Amita
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