[Histonet] ATPase 9.4 Troubleshooting

[Bryan, Karen]

Dear Friends,

I am having trouble with the ATPase 9.4 stain on muscle biopsies performed on frozen sections.
The sections look extremely dark and almost have a digested look to them.

To troubleshoot we:


*         made all new solutions,

*         opened a new container of ATPase (stored in the -15 deg. C. freezer)

*         the oven has been tested and confirmed to be stable at 40 deg. C.

*         the pH is stable at 9.4 (+/- 0.02)

*         only distilled water is used for all solutions where required and for rinses between steps.

The ATPase 4.3 has some artifact but definitely is readable and not like the 9.4. All of the other sections for the other stains in the protocol (H&E, Trichrome, NADH, ORO, PAS) are coming out great and the sections do not show any freezing or microtomy artifacts.

Has anyone else run into this problem? We have run the same muscle cases 3 times in the last 2 days after making the changes to all of the solutions to prepare the ATPase.

What are we missing? I may be able to upload a photo if you need to see the issue clearly.

Thank you so much for your help!

Karen

Karen P. Bryan, HT (ASCP)cm
Sr. Histology Specialist
Department of Pathology/Amory 3
Brigham & Women's Hospital
75 Francis Street/Boston
617-732-5454
(Internal extension 2-5454)



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