Thanks for the quick response. I would love to take a look at your method. Our method from Dubowitz's 2nd edition has worked fine for years, but can leave residue on the slides, especially when fresh, fades & is very messy to file and we seem to be going back to previous bx's with more frequency. The 3rd edition of Dubowitz & Dawson's Neuropathology Techniques do not use PVP or glycogen, but use Dextran. This solution is much nicer to work with, not as thick & does not leave deposits on the slide when fresh and claims to appear stable for several years. So, finally, after doing this test one way "because we have always done it that way", I am trying to clean it up a little. Also, do you do phosphorylase routinely, if not what is the criteria for doing it. At present we do it as a part of a panel for all muscle bx's (which amounts to 80 to 110 per year). It seems this may not be necessary. Thanks again for your interest; I would appreciate your input. Have a good day, Sharon Allen
Senior Medical Technologist Neuropathology Lab-MS435U Health Sciences Centre 820 Sherbrook Street Winnipeg,MB, CA R3A 1R9 e-mail: sal...@dsmanitoba.ca -----Original Message----- From: Mitchell Jean A [mailto:jmitch...@uwhealth.org] Sent: Wednesday, March 28, 2012 9:07 AM To: Sharon Allen Subject: RE: [Histonet] Phosphorylase method Sharon: Just curious - Does the phosphorylase method you use fade after time? The method I use does use PVP and dextran (no glyceron though). I am not certain of why these compounds are used; may have something to do with carbohydrate bonding but wouldn't swear by it. I make the incubation media up in larger batches; aliquot into smaller amounts; freeze at -20 until ready to use. I section my muscle onto 22 X 22 coverslips and only use 10ml aliquots of incubation media at a time. Your name sounds familiar; possibly from one of the muscle workshops I have given at NSH? Anyway - if you don't already have my procedure - let me know and I will forward. Jean Mitchell, BS HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Manager 600 Highland Avenue Madison, WI 53792-5132 -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Sharon Allen Sent: Wednesday, March 28, 2012 8:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Phosphorylase method Hi, Does anyone have a good enzyme method for Phosphorylase done on muscle bx's. We have been doing the method from Dubowitz, 2nd edition for years but would like to get away from the messy PVP & coverslipping. We are trying cytoseal, which seems to work fine for coverslipping, but would like to compare other methods without PVP. Also, does anyone know the purpose of PVP, glyceron or dextrin in this test? Thanks for any help, Sharon Allen Senior Medical Technologist Neuropathology Lab-MS435U Health Sciences Centre 820 Sherbrook Street Winnipeg,MB, CA R3A 1R9 e-mail: sal...@dsmanitoba.ca
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