Several ideas - 1. Make sure the biopsies do not dry before adding to the fixative. 2. Extend your fixation time as much as possible, decrease your processing times (esp if you are using microwave processing) 3. Dry the slides for 25-30 min at 64oC (60 may not be hot enough to melt the paraffin
Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Emily Butte Sent: Friday, 31 July 2009 11:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Washing Our lab has been struggling for a while now, with tissue washing off the slides. The problem was originally traced to our processor that was malfunctioning and having major carry-over issues, causing poor tissue processing. So after failed attempts to get the old machine fixed, we ended up getting a new microwave processor. The tissue (mainly prostate bx) comes out of the processor looking good. It cuts well, but after staining, some of the sections are no longer on the slides. The problem was quiet severe with the old processor and has decreased drastically, but has not entirely gone away with the microwave processor. Here's some info on our current procedure. This is a small physician office lab and we everything manual. We use plus slides and a water bath with no tissue adhesive added. The bath is between 42-45 degrees Celsius. After sectioning, the slide are put in an oven at 60 degrees for 20 minutes. After taking the slides out of the oven, we cool them to room temp then hand stain them in an H&E line. Xylene - 5 min Xylene - 2 min Xylene - 2 min 100% Alcohol - 30 sec 100% Alcohol - 30 sec 100% Alcohol - 30 sec 95% Alcohol - 30 sec Rinse in DiH2O Hematoxylin - 2 min Rinse in DiH2O 0.50% Acid Alcohol - 2 dips Rinse in DiH2O Scott's Tap Water - 10 sec Rinse in DiH2O 95% Alcohol - 30 sec Eosin - 30 sec 95% Alcohol - 30 sec 100% Alcohol - 30 secs 100% Alcohol - 30 secs 100% Alcohol - 30 secs Xylene - 30 sec Xylene 2 min Xylene - 2 min The slides are then cover slipped and of a 12 piece prostate case, at least a section or 2 will wash off of about 4 of the slides. Please pass along any ideas as to what the cause of this could be and how I can fix it. I'm fresh out of ideas and would really appreciate the advice of someone with more experience than myself. Thanks, -- Emily Path Lab Solutions Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
