[Histonet] (no subject)

[Tony Henwood (SCHN)]

The reason for using diastase is to remove the glycogen from the tissues. AND if you do not heat the methenamine silver solution for the GMS it will take forever to stain, in fact all you will get is background spots. This I can verify, have you ever forgot to turn on the water bath before staining? - several hours later - nuthin!!

Who ever is giving you the advice I would wonder about their histotechnology education and training (in fact have they had any training at all?).

Do not listen to them. Extrapolate from cytology. GMS and DiPAS have been done on cytology smears fixed in ethanol as well as air-dried for years see:

Loughman NT. "Pneumocystis carinii: rapid diagnosis with the microwave oven" Acta Cytologica. 33(3):416-7, 1989 May-Jun.

PINTOZZI, R. L. "Modified Grocott's methenamine silver nitrate method for quick staining of Pneumocystis carinii"  J Clin Pathol 1978 31: 803-805

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ingles Claire 
Sent: Wednesday, 15 February 2012 6:22 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] (no subject)

Hey histo gurus:
Just a little twist for you. We are planning on doing GMS and PASD on frozen skin sections. Is there any change in staining protocol for paraffin vs frozen. We plan on fixing in ETOH. I have a co-worker saying that we don't need to use diastase on the PASD because there are already active "enzymes" in the tissue as it is fresh. He also said that we don't need to heat the silver for the GMS. Not sure why. These changes don't seem right to me. But I have been wrong before (Yes, I admit it). Thanks for all the wisdom out there floating in the electrons. We are staining for fungus BTW.
Claire

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