[Histonet] Bone decalcifying/processing problem

Elizabeth Chlipala liz <@t> premierlab.com
Fri Jul 12 09:55:38 CDT 2013


I know this might sound silly but normal mouse bone marrow contains both fat and hemoatopoietic marrow,  the areas that are fat will appear like round voids.  Is that what you are seeing?


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

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From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Orla M Gallagher [o.m.gallagher <@t> sheffield.ac.uk]
Sent: Friday, July 12, 2013 8:52 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Bone decalcifying/processing problem

Dear Histonetters,

Has anyone ever seen large holes/vacuoles in the bone marrow of mouse
long bones after decalcification and processing to wax? We've had this
problem with a study which was fixed in 4% paraformaldehyde and
decalcified at 4 degrees C over a 14 day period according to a
routinely-used protocol involving changes of 15% EDTA/0.5% PFA in PBS
pH 8.0, rinsed in PBS to remove EDTA before processing to paraffin
wax. The marrow is filled with large holes, almost like balloons. I'm
happy to send images to anyone who might be able to help!

Best wishes,

Ms. Orla Gallagher
Bone Analysis Laboratory
Mellanby Centre for Bone Research
Department of Human Metabolism
D Floor Medical School
University of Sheffield
Beech Hill Road
S10 2RX

Website: http://mellanbycentre.dept.shef.ac.uk

Tel:         0044114-2713337 (office)
              0044114-2713174 (lab)
E-Mail:    o.m.gallagher <@t> sheffield.ac.uk

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