[Histonet] mouse ankles for IHC

[Dana Marshall]

Hi everyone,

I am gearing up for mouse ankle-joint histology.  (I may do vertebral column eventually as well if anyone has done that.)  I have a Cryojane tape transfer system and have done a bit of sectioning of the undecalcified bones and they look adequate.  I haven't done any fancy staining yet.  My question is a fairly broad one in that I wanted to ask those who might be doing this for a brief outline of their procedure from beginning to end.  So far I am fixing in 10% formalin (in H2O) for a couple of days.  I embed the joint with the heel towards the bottom of the block (I thought it might be more stable to cut up from the heel but perhaps not?)  Ultimately I will do IHC staining and, possibly, some in situ hybridization.  I have some previous info in regards to knives and angles of cutting etc and would appreciate any additional insights into that as well.

I am having trouble stringing together the information that will allow me to make wise choices early on to maximize the use of my samples (and my mice).  Any advice would be GREATLY appreciated.

Thanks again.

Dana (who dabbles in histology every 10 years whether I need to or not:-)