[Histonet] does anyone know how to get better immersion fixation of mouse femur?

[Gayle Callis]

Sharon,

If you do not need the middle of the femur, or diaphysis, and only 
interested in the two ends where majority of marrow resides in femur,  then 
cut the bone open in middle to permit internal access for fixative from 
these open ends.   If possible, try to get a minibone saw for this, i.e. 
Dremel tool or similar tool, so you don't crush the bone.  Poking holes in 
mouse femurs is more likely to destroy the marrow you want to see since the 
bone shaft is so tiny - just achieving a tiny hole may be difficult, 
although this is a technic that works for large animal bones - cutting 
windows or drilling holes away from lesions or areas you need to see.

   The other alternative it to open both ends of the femur and push the 
bone marrow out.  One lab here does that for mouse bone marrow and is fine 
IF you do not want to see trabecular bone with the marrow, cortical bone or 
cartilage.   This is more like a bone marrow biopsy.

At 09:19 AM 3/9/2007, you wrote:
>Dear Histonetters,
>
>I have been using fixed mouse femur to do mouse bone marrows.  I get much 
>better fixation and preservation of bone marrow if I perfuse the mouse 
>than if I immersion fix the femurs.  However, I prefer not to perfusion 
>fix the mouse because perfusion fixation prevents me from collecting other 
>tissues unfixed.  Is there a way to improve the fixation of the mouse 
>femurs when I immersion fix?  I'm guessing that the problem is I'm not 
>getting good enough penetration - I suppose I could poke a few holes in 
>the femurs?
>
>Thanks in advance for your help,
>Sharon
>
>_______________________________________________
>Histonet mailing list
>Histonet <@t> lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)