[Histonet] Paraformaldehyde Fixed Tissue

[Ana Maluenda]

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.Maluenda at baker.edu.au W www.baker.edu.au

-----Original Message-----
From: Sandra Cheasty [mailto:sandra.cheasty at wisc.edu]
Sent: Tuesday, 29 August 2017 1:30 AM
To: Histonet (histonet at lists.utsouthwestern.edu) <histonet at 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


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