[Histonet] RE: Mouse bone blocks/sections, bone marrow smears

[Elizabeth Chlipala]

Jian

I would fix for about 48 hours, then decal overnight in 10% formic acid, process to paraffin on a cycle of around 45 minutes per station, we sometimes process on hour long cycles with 3 absolutes and 3 xylenes, the sections cut just fine.  We trim and then then soak the blocks for about 30 minutes on wet ice then section at 5 microns, let the slides drain a bit, shake off excess water and lay flat on a hot plate overnight set at 40C.   If you are preparing bone marrow smears you want to keep then away from any 10% NBF so if you are collecting at necropsy stay as far away from the formalin as possible that will mess up your stain.  Crack the femur and with a paint brush dipped in 5% EDTA collect the bone marrow from the femur and then "paint" it on the slide.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Laboratory Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
Work (303) 682-3949
Fax (303) 682-9060
Cell (303) 881-0763
liz <@t> premierlab.com
www.premierlab.com

Ship to address:

1567 Skyway Drive, Unit E
Longmont, CO 80504


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Yu, Jian
Sent: Thursday, March 21, 2013 5:27 PM
To: 'histonet <@t> lists.utsouthwestern.edu'
Subject: [Histonet] Mouse bone blocks/sections, bone marrow smears

Dear all,

We like to make paraffin embedded bone of mouse tibia and cut longitudinal sections for H&E sections.

Does anyone have a easy-to-follow protocol to prepare (fix and decal) the bone before processing? We routinely fix soft tissue (ie. mouse GI tract  and liver) and send off to our pathology core for processing and embedding.

Any tips on cutting the blocks is helpful too. These sections appear brittle.

We are also interested in a practical protocol for making mouse BM smears and staining to determine cellularity changes after radiation.

We just started working with the bone.  Any advice will be very helpful and greatly appreciated.

Thanks, Jian
*******************************************************************
Jian Yu, Ph.D.
University of Pittsburgh Cancer Institute
Email: yuj2 <@t> upmc.edu
*******************************************************************


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