Dear All, 

Isn't glycogen water soluble? You may need to use different fixative to
retain glycogen in the first place, Hrapchak and Sheehan book has a list of
fixatives recommended for different tissue components. Carnoys or alcoholic
formalin, even pure alcohol fixation is recommended for glycogen storage
disease with aqueous fixatives avoided.  Hence NBF may NOT be the fixative
of choice for what you want to see. We found this with fish liver project
and if you have a research project going, a piece of liver in the proper
fixative may solve some of your problems with other pieces collected in NBF
for routine staining, etc.   It very well may NOT be the staining method,
but initial handling. This way the experiment is controlled with the
special stain on THE target tissue (liver) containing glycogen.  


At 08:52 AM 1/16/2004 -0600, you wrote:
>Thank you all for your ideas, let me elaborate...
> 
>Our pathologists won't except cervix for a control, they want a liver
>control. When we use cervix, the stain seems to work great.  I have not
>tried skin or kidney, but maybe I can get them to except them as controls.
>We have tried spitting, it works best.  I don't feel it is a problem with
>our procedure, because it seems to work well on cervix and the Schiff's
>reaction seems to be very strong.  Freida, you may have hit the nail on the
>head with your comment on glycogen storage in the liver.  Autopsy tissue is
>about all the liver we can get.  Could be that we will never get good,
>consistent results with liver controls.  However, in my previous lab we did
>get consistent staining with liver.  That is what has me so perplexed.  I
>think I will try the kidney.  If alot of you respond that liver will never
>make a good control, maybe I can use that as proof to the pathologists that
>we need to use the cervix control.  Any other ideas....please let me know.
> 
> 
>[Barnhart, Tammy]  -----Original Message-----
>From: FreidaC@aol.com [mailto:FreidaC@aol.com]
>Sent: Friday, January 16, 2004 8:21 AM
>To: Tbarnhart@primecare.org; histonet@lists.utsouthwestern.edu
>Subject: Re: [Histonet] PAS with and without Diastase for glycogen
>
>
>
>Tammy:
> 
>Many livers do not contain a lot of glycogen - especially autopsy livers,
>because few are well nourished and have a lot of stored glycogen at the time
>of death.  A better control in my opinion is a section of cervix containing
>both endo- and ecto- cervix.  The stratified squamous epithelium of the
>ectocervix contains glycogen which will disappear with diastase digestion,
>while the mucous glands in the endocervix do not contain glycogen, but
>rather mucin that will be positive with the PAS.  This will not disappear
>with diastase digestion.  This is also easier control tissue to find in most
>institutions.
> 
>Freida Carson
>
>
>
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology 
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)



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