Hi Julie, Try increasing your deparaffinization times, sometimes there is just not enough time in xylene so subsequent staining can be very inconsistent like described.
Kari Kienitz HT, (ASCP) Histology Laboratory Gastroenterology-EAST The Oregon Clinic 1111 NE 99th Ave Portland, OR 97220 503.935.8311 kkien...@orclinic.com CONFIDENTIALITY WARNING: This e-mail and any attachments are for the exclusive and confidential use of the intended recipient. If you are not the intended recipient, please do not read, distribute or take action in reliance upon this missive. If you have received this in error, please notify the sender immediately by reply e-mail and delete this message and its attachments from your computer system. Thank you ________________________________________ From: histonet-boun...@lists.utsouthwestern.edu [histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Julie Cohen [juc2...@med.cornell.edu] Sent: Wednesday, April 01, 2015 9:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] inconsistent H&E staining Hi, I made slides of paraffin-embedded mouse small intestine (Swiss rolls), and stained them with Hematoxylin and Eosin. Parts of the tissue on the same slide are stained dark with good structure. Other areas look washed out with poor structure. We realize that some of this could be caused by the orientation/structures captured, but similar tissue type looks paler as well. Has anyone had a similar experience, and could suggest an explanation for me to give to our client? At first we thought it might be due to poor fixation, since the centers of tightly-wound rolls were affected, but we also observed this in the outer parts of loosely wound rolls. I soak the blocks before sectioning; could non-uniform swelling result in variations of the section thickness? (These are 7 microns thick.) Apologies if this information is available somewhere else; I tried unsuccessfully searching the archives. Thank you, Julie Cohen Research Lab Tech EM Core Facility Weill Cornell Medical College 1300 York Avenue, Room A-105 New York, NY 10021 lab: 212-746-6146 email: juc2...@med.cornell.edu<mailto:juc2...@med.cornell.edu> _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
