[Histonet] PAS with Glycogen Digestion - Solution Not Working

[pam johnson]

What is wrong with saliva. I have done this procedure for 20 years. It works beautifully.


-----Original Message-----
From: Mickie Johnson <mickie25 <@t> netzero.net>
To: rjbuesa <@t> yahoo.com; histonet <@t> lists.utsouthwestern.edu; 'McKnight, Tanisha' <tanisha.mcknight <@t> covance.com>
Sent: Sun, 26 Oct 2008 9:09 am
Subject: RE: [Histonet] PAS with Glycogen Digestion - Solution Not Working



Diastase powder will dissolve in any water, but requires some calcium ions
o function. Thus it will not perform in distilled water, but will perform
ell using plain tap water. Also, the diastase powder from sigma will work
ell if only a very small amount of the powder is used. We always used just
nough to barely cover the bottom of a glass coplin jar and then fill with
ap water. I would expect 0.5 grams in 50 mls of water is probably too much.
ry using one tenth that amount or 0.05 gm in 50 ml. When there is too much
iastase in solution, enzyme conformation is inhibited and it  does not
ork. Spokane water is quite hard. It always worked well at room temperature
or 20 minutes.
Mickie

ickie Johnson, B.S., HTL(ASCP)
ohs Histology Consulting Services, LLC
 & Mohs Lab Staffing
507 S. Manito Blvd.
pokane, WA 99203
09-954-7134
AX   509-624-3926
eb: www.mohshistogyconsulting.com & www.mohslabstaffing.com 
mail: mickie25 <@t> netzero.net

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----Original Message-----
rom: histonet-bounces <@t> lists.utsouthwestern.edu
mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
ent: Friday, October 24, 2008 8:38 AM
o: histonet <@t> lists.utsouthwestern.edu; McKnight, Tanisha
ubject: Re: [Histonet] PAS with Glycogen Digestion - Solution Not Working
Tanisha:
eing an enzyme, you cannot use diastase in distilled water. You have to
repare a diastase buffer at pH7 to dissolve the diastase and do the
igestion at 37ºC during 30 minutes. That will work.
lease, do not try to use saliva!
ené J.
--- On Fri, 10/24/08, McKnight, Tanisha <tanisha.mcknight <@t> covance.com>
rote:
From: McKnight, Tanisha <tanisha.mcknight <@t> covance.com>
ubject: [Histonet] PAS with Glycogen Digestion - Solution Not Working
o: histonet <@t> lists.utsouthwestern.edu
ate: Friday, October 24, 2008, 11:26 AM
Hi All:

 am in the process of working up a PAS Staining protocol. The PAS stain
s working fine, but I am unable to get my tissue to digest. I am using
5g of Diastase in 50ml of DI water. I tried heating the solution in a
ater bath for 30 min, 1 hour and 1.5 hours and it still looks the same
s the one without digestion. Could I be using the wrong Diastase? If
o, does anyone have any suggestions for a new product?

hanks,

anisha Neely, HT(ASCP)
P-Histology/Specimen Management
<mailto:tanisha.mcknight <@t> covance.com>  

 

     
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