Hi Sandra, I haven't myself particularly worked with brain and spinal cord, but majority of my protocols in my old job used fixation in 4% PFA (24 hours at 4-8oC) and routinely process (or transfer to graded EtOH, if not processing immediately). However, our routine process didn't include a first step in 10% NBF, since PFA plays the role of fixation. Therefore, after PFA, we would have 70% EtOH, 80% EtOH, 95% ETOH, 2x EtOH the Xylenes and wax [assuming you are referring to FFPE?]. Also mouse tissue can be small and delicate, so I remember running liver, spleen, kidney and thymus (soft tissues I worked with) in a short cycle (similar to what I would do for biopsies).
Hope this helps! Kind regards, Ana Ana Maluenda Research Assistant Atherothrombosis and Vascular Biology Laboratory Baker Heart and Diabetes Institute 75 Commercial Road, Melbourne VIC 3004 P (03) 8532 1359 E ana.malue...@baker.edu.au W www.baker.edu.au -----Original Message----- From: Sandra Cheasty [mailto:sandra.chea...@wisc.edu] Sent: Tuesday, 29 August 2017 1:30 AM To: Histonet (histonet@lists.utsouthwestern.edu) <histonet@lists.utsouthwestern.edu> Subject: [Histonet] Paraformaldehyde Fixed Tissue Hi all, We are having difficulty sectioning mouse tissue, (brain, spinal cord, liver, and spleen), on paraformaldehyde fixed tissue. Has anyone had issues with paraformaldehyde fixed tissue? They were processed routinely, starting in 10% NBF, with other tissues, and we are cutting them at 3u. Thank you! Sandy Sandra J. Cheasty, HT (ASCP) Histology & Necropsy Supervisor UW-Madison, School of Veterinary Medicine Protecting your privacy is important to us. The Baker Heart and Diabetes Institute will handle your information in accordance with the Privacy Act 1988 (Cth) and its Privacy Policy which is available at www.baker.edu.au or on request by contacting priv...@baker.edu.au or by calling 1800 838 498. The Privacy Policy also explains how you can access and correct your personal information, or make a complaint about a breach of the Australian Privacy Principles. bidipp2014.0.1a -- Message protected by MailGuard: e-mail anti-virus, anti-spam and content filtering.http://www.mailguard.com.au/mg _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
