Sowmya:

Please clarify.  What kind of histomorphometry must you do that requires decalcification.  Distance between labels?  That is extremely easy on PMMA embedded thin sections.

Tim

I will have to perform growth plate cartilage histomorphometry, immunohistochemistry for various antibodies. 

OK, that explains it then.  The growth plate cartilage histomorphometry is straightforward on PMMA-embedded thin sections (there are great surface staining routines), but while there are some immunohistochemistry protocols for PMMA, they are not for many, and they are fickle.  We have had limited success by decalicifying the surface mineral and applying immunohistochemistry stains to the surface.  You could try that, which will allow you to have direct correlative microscopy with your calcein label.  Alternatively, you could divide your sample in half, one for PMMA calcified and one for decalcified work.  If you can't do that, and must embedd all first, then you can later remove the plastic with heptane or chloroform.

Tim

 Sowmya, Even though you can get ride of all PMMA from your bone, the tissue's antigenicity has damaged (or partially damaged ). Maybe it can be done on non-immunostaining. 

First, I am sure you do not have to decalcify the tissues to do histomorphometry.  You an do this directly o the PMMA embeded samples as Tim Bromage indicated.

In terms of reusing PMMA embedded calcified tissue; as Dorothy Hu pointed out it depends on exactly what you want to measure.  I am pretty sure that you can remove the PMMA but the damage you do to the tissues during this process may limit what you can do after that.

Hi Sowmya, we do decalcify sections regularly (before TRAcP staining) using 2-methoxyethyl acetate (MEA), and this works quite well. We have made some antibodies work on tissue prepared this way, but it required complete optimization of immunostain conditions for each, and quite a few antibodies we just could not get to work. In my experience it is better to use one bit of tissue (for instance left leg) for mma and another (right leg) for wax or cryosections.

 Hi Sowmya, calcein labelled tissue (bone and cartilage) can be included in PMMA resin for histomorphometry using routine procedures

i hope thise help you.

Sowmya -If you decalcify the bones you will remove all of the calcein label so I would recommend that you do all of your histomorphometry on the PMMA embedded sections first.  It is a long procedure to redissolve the plastic, decalcify the bone and reprocess in paraffin and, as Dorothy indicated, most likely your antigens will suffer damage through all of this processing.  We have had limited success staining MMAembedded bone  via IHC for antigens such as collagen I, II, osteopontin and osteocalcin, but have not had any success staining for antigens such as CD markers.