Not only heat affects IHC results, especially on bone marrow biopsies; decalcification also can greatly affect your results.  Formic acid decalcification is recommended if you want to do IHC on bone marrow biopsies.

If you have a particle prep or clot section, I'd certainly try the IHC on those to see if you can get a better result.  It is also possible that the antibody does not react in 100% of the cases.

Teri Johnson
Managing Director Histology Facility
Stowers Institute for Medical Research
1000 E. 50th St.
Kansas City, Missouri  64110
tjj@stowers-institute.org

-----Original Message-----
From: Kim Saunders [mailto:cactusmoon782000@yahoo.com]
Sent: Friday, December 27, 2002 3:20 PM
To: histonet@pathology.swmed.edu
Subject: DBA44


Hi,
I'm trying to work up the DBA44 (Dako) on the Ventana ES IHC Stainer. I am currently trying the recommended 1:25 dilution for 32 minutes. I've tried our NordicWare pressure cooker Antigen Retrieval method (20 in the microwave, cool down for 20 minutes), and Protease 1 for 4 & 8 minutes, and Protease 2 for 8, 12, 16, & 20 minutes. I have a bone marrow bx that the pathologist is sure should be positive based on the flow results. Since my ES uses heat, could the heat be an issue? Any help would be greatly appreciated.
Thank you,
Kim Mann-Saunders
St. Joseph Hospital-Tucson, AZ


Kim Saunders
Proud to be an American!!




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