H&E of decalcified bone samples

From:"Kinsley, David" <david.kinsley@spcorp.com>

Hi histoneters,
I was hoping someone could help me out with a staining question.  I process
mouse tissues and am currently working with front and hind paws and joints
to study inflammation.  I use Cal rite from Richard Allen and decalcify
according to their recommendations.  Usually no more than 24-48 hours for
hind limbs.  (This is after 4-5 days of fixation in NBF) I also perform all
fixation and decal on a tissue shaker to help facilitate penetration of
reagents.  I feel this is the minimum to have demineralized samples without
jeopardizing staining quality.  However, there is more eosinophilic staining
occurring thank undec'd samples from the same animals.  I know that this is
a result of the acid decalcifying process(and we don't have time to use
EDTA)   Is there any way of soaking the slides in a higher pH PBS to bring
the samples to pre-decal Ph to improve nuclear staining?  Any other
suggestions are appreciated.  The staining is throwing off the ability to
interpret which cell types are in the tissue.

Thanks in advance

Dave Kinsley MS,HT/HTL(ASCP) 

Schering-Plough Research Institute
Department of Immunology 
2015 Galloping Hill Road
Kenilworth, New Jersey 07033

Phone:  908-740-4669
Fax:      908-740-3084

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