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) The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet