[Histonet] Bone decalcifying/processing problem

Orla M Gallagher o.m.gallagher <@t> sheffield.ac.uk
Fri Jul 12 09:52:03 CDT 2013

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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