[Histonet] Re: Ammonia water pH and other bluing solutions

[Rittman, Barry]

When I was training (I admit this was soon after Noah landed the ark),
we used tap water to blue sections. This was in London and the water was
slightly alkaline at that time.
Blueing solutions should be used with great care. 
Most blueing solutions give essentially the same result. 
There has been a lot of discussion re using lithium carbonate and
ammonia solutions, however very little discussion regarding the
dilutions at which these should be used. The smallest amount of a
"blueing" agent that will result in blueing of the hematoxylin should be
used. It should then be carefully washed to remove all traces. 
If the solution is too strong it can remove some of the hematoxylin,
tend to lift the section off the slide and if not washed out carefully
will interfere with the binding of eosin.
Barry

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kathleen
Roberts
Sent: Wednesday, December 01, 2004 8:35 AM
To: Jackie M O'Connor; 'histonet <@t> lists.utsouthwestern.edu'
Subject: Re: [Histonet] Re: Ammonia water pH and other bluing solutions

Heck, we still use that.  All our slides look lovely.

The mellow one-
Kathleen Roberts
Rutgers University

Jackie M O'Connor wrote:

>Anyone remember using saturated lithium carbonate?  I could probably
use a 
>little lithium right now . . . . . 
>
>Correct me if I'm wrong (Pandora's box) but isn't the "bluing" step
just 
>bringing the slides BACK to a neutral pH after treating them with acid 
>which makes them purple-ish?  I like the ammonia because it 'seems' to 
>make the nuclei sharper and instantaneously - Li2Co3 and PBS seemed to 
>make them "blah".(Perception is in the eye of the beholder)   Running
tap 
>water will make the same miracle happen, but it takes longer.
>
>
>
>
>
>
>"Johnson, Teri" <TJJ <@t> Stowers-Institute.org>
>Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
>11/30/2004 12:51 PM
>
> 
>        To:     "Histonet" <histonet <@t> lists.utsouthwestern.edu>
>        cc: 
>        Subject:        [Histonet] Re: Ammonia water pH and other
bluing solutions
>
>
>Way back in the day, Brigati was using PBS on his immunostainer to blue
>after the hematoxylin counterstain.  I have since used PBS to blue my
>slides instead of ammonium hydroxide/water.  It's cheap, and very easy
>on the sections, and I have plenty of it on hand.
> 
>
>Teri Johnson
>Managing Director Histology Facility
>Stowers Institute for Medical Research
>1000 E. 50th St.
>Kansas City, Missouri  64110
>tjj <@t> stowers-institute.org
>
>
> 
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