What is wrong with saliva. I have done this procedure for 20 years. It works beautifully.
-----Original Message----- From: Mickie Johnson <[EMAIL PROTECTED]> To: [EMAIL PROTECTED]; histonet@lists.utsouthwestern.edu; 'McKnight, Tanisha' <[EMAIL PROTECTED]> 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: [EMAIL PROTECTED] ISCLAIMER: his message is intended for the sole use of the addressee, and may contain nformation that is privileged, confidential and exempt from disclosure nder applicable law. If you are not the addressee you are hereby notified hat you may not use, copy, disclose, or distribute to anyone the message or ny information contained in the message. If you have received this message n error, please immediately advise the sender by reply email and delete his message. ----Original Message----- rom: [EMAIL PROTECTED] mailto:[EMAIL PROTECTED] On Behalf Of Rene J Buesa ent: Friday, October 24, 2008 8:38 AM o: histonet@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 <[EMAIL PROTECTED]> rote: From: McKnight, Tanisha <[EMAIL PROTECTED]> ubject: [Histonet] PAS with Glycogen Digestion - Solution Not Working o: histonet@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:[EMAIL PROTECTED]> ______________________________________________ istonet mailing list [EMAIL PROTECTED] ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet No virus found in this incoming message. hecked by AVG - http://www.avg.com ersion: 8.0.175 / Virus Database: 270.8.3/1747 - Release Date: 10/26/2008 :27 AM ______________________________________________ istonet mailing list [EMAIL PROTECTED] ttp://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet