Cynthia, Hematoxylin will inactivate the peroxidase (link) in the tissues. You must go back and re-do the link step and hopefully the antigenic sites will still accept more streptavidin (or similar) conjugated antibody. Cheer. Greg Greg Dobbin, R.T. Chief Technologist, Anatomic Pathology Dept. of Laboratory Medicine, Queen Elizabeth Hospital, P.O. Box 6600 Charlottetown, PE C1A 8T5 Phone: (902) 894-2337 Fax: (902) 894-2385 "I find that the harder I work, the more luck I seem to have." - Thomas Jefferson
>>> "Cynthia Pyse" <cp...@x-celllab.com> 1/24/2011 8:57 AM >>> Happy Monday Histonetters I need to pick the brains of the IHC experts out there in histoland. I run IHC on the Dako autostainer. On one of our runs there was not enough DAB to dispense on all the slides. My tech dehydrated the slides and cover slipped them. We printed the stain log on the run and confirmed that the DAB was not dispensed on three slides. The cover slips were removed and the slide were soaked in xylene to remove the cover slipping media. The slides were rehydrated and DAB was applied. There was still no reaction after 10 minutes, The DAB was reapplied for up to 30 minutes with no reaction. Can anyone out there give me a reason that the slides would not react after the reapplication of the DAB. I have done this before when the DAB solution was short on the run and it worked. I can't figure out why this didn't react. I called tech service at Dako and talked to the supervisor, she agrees with me that the reapplication should have worked. I checked the run log and the only missing step is the DAB. The antibodies with the missing DAB have their own controls on the slides. Any thought out there as to what the problem is. I have had the PM done on the Dako which should resolve the problem according to Dako. There has not been a problem since. This mystery will drive me crazy until I can figure out why. I could use help. Thanks in advance, I appreciate any input. Cindy Cindy Pyse, CLT, HT (ASCP) Laboratory/Histology Supervisor X-Cell Laboratories 716-250-9235 e-mail cp...@x-celllab.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------- Statement of Confidentiality This message (including attachments) may contain confidential or privileged information intended for a specific individual or organization. If you have received this communication in error, please notify the sender immediately. If you are not the intended recipient, you are not authorized to use, disclose, distribute, copy, print or rely on this email, and should promptly delete this email from your entire computer system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
