For the pale H&E, has the wax been completely removed in the xylene. If slides are dried upright, then the wax flows down the slide possibly covering the last section, requiring longer in the dewax solution. Secondly, after the differentiation stage, the acid/alcohol again may be collecting at the bottom of the slide especially in the automatic stainers prior to water rinsing (increasing the differentiation time for the lowest section). To differentiate these two causes, is the eosin pale as well? If it is then it might be caused by the incomplete de-waxing.
For the long NBF fixation of placenta, has excess blood been removed from the placenta? Or have you checked the concentration of formalin (see Jaspers (1987) J Histotechnol 10 (4): 263-265 for the method using Schiffs reagent). 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: [EMAIL PROTECTED] [mailto:[EMAIL PROTECTED] On Behalf Of Karla Arrington Sent: Wednesday, 19 November 2008 12:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Uneven H&E Staining Recently we have had an issue with uneven H&E staining. Lately slides that have three sections on one slide, mostly GI biopsies, and the last section (bottom of slide) are paler than the upper two section. All cut by the same person and the stains were great. What could be the cause of this? Also with the quality of formalin. Can formalin be purchased as "bad" when it takes 6 days to fix placenta or breasts? What is the cause? Karla Arrington, HT(ASCP) _______________________________________________ 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
