[Histonet] RE: Decalcifying and Processing large bones

[Cesar Francisco Romero]

For many years now I have used this protocol in Bone
Marrow samples.

The cortical of the bone is still a problem, but it
can be solved with longer decal time.

If you need to preserve morphology and antigenicity
for Immunohistochemistry this is the best choice.

Three Days Bouin’s Fixative ( From the very Beginning
), don’t use Formalin.

Three Days of a Saturated Solution of EDTA ( Disodium
Salt ) in distilled water.

After this time put the blocks in the first alcohol and
process as usual.

Don’t put the blocks in formalin.

EDTA alone or with Heating  affects morphology very badly.

For IHC Antigen Retrieval is the recommended or used
for each antibody.

Difficult ones as CD 5 or CD 23 work very well with
this decal protocol.

Cesar
Romero

Buenos
Aires.

Argentina.