[Histonet] Perfusion of mouse bone femur

[Reuel Cornelia]

Does anybody have a Protocol for perfusion of mouse bone femur. It looks like my protocol does not work.
1.fix in 4% PFA in PBS for 24 hrs in refrigerator
2. wash in PBS 3x 
3. transfer in 14 %EDTA for 5 days.
4. Wash in water for maximum of 2 hrs in running water
6. perfusion with 15% sucrose overnight
7.perfusion with 30% overnight.
8. Transfer in OCT.
8. Cut in 10 um and transfer section on APES slide
 
Results: When cutting section, the tissue does not adhere to the slides properly.
 
Can you please help us or any comment to correct this protocol.
 
 
Reuel Cornelia, BS MT, AMT
Cellular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
Tel: 214-559-7766
fax: 214-559-7768