Hi Karen- I have also had problems getting tissue being stained for IHC markers to adhere to the slide. The Potassium permanganate is really harse on the tissue (if you can get the sections to stay on). Here is a trick I picked up at an NSH lecture that I have used successfully several times.
Run your IHC stains as usual. Rinse well in water. Do not counterstain with hematoxylin. Stain with DiffQuik II for 30 seconds. Rinse in water for a short period of time. Differentiate with 10 dips each in 2 changes of 95% ETOH and 2 changes of 100% ETOH and then clear in xylene and coverslip. Results: melanin pigment will turn green. Brown DAB stain will be unaffected. Other tissue elements will be stained a medium blue color. This method will also work for other Special Stains but I have not attempted to modify this method for use on "Red" stains. Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA. -----Original Message----- From: Heckford, Karen - SMMC-SF via Histonet [mailto:[email protected]] Sent: Tuesday, May 10, 2016 8:45 AM To: [email protected] Subject: [Histonet] Melanin Bleach Good Morning, I need some help. Yesterday I bleached some heavily pigmented tissue. I have to run some IHC's on them. I bought the bleaching kit from American Master Tech. I had to put the Permanganate for about 6 hours to get the melanin and then a couple of minutes in Oxalic Acid. I had to let them set overnight because I could not get another IHC run in that day. It looks like the tissue fell off during decloaking. I used Apex slides. I rarely ever have to bleach anything here. So I am not sure if I did this correctly. I am thinking I need to bleach and do the run in the same day and not let them set over night in DiH20. Thanks, Karen Heckford HT ASCP CE Lead Histology Technician St. Mary's Medical Center 450 Stanyan St. San Francisco, Ca. 94117 415-668-1000 ext. 6167 [email protected] Caution: This email message, including all content and attachments, is CONFIDENTIAL and may be of a nature that is LEGALLY PRIVILEGED. The information contained in this email message is intended only for the use of the recipient(s) named above. If the reader of this message is not the intended recipient or an agent responsible for delivering it to the intended recipient, you have received this document in error. Any further review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please notify us immediately by reply email. Thank you." _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet This email and attachments may contain PHI that is privileged and confidential and is not intended for any unauthorized person. If you, the reader, are not the intended recipient you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. Do not read the email but instead reply to the sender and destroy the message and any attachments. Thank you. _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
