Re: Toluidine blue stain for undecalified bone (MMA)

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Dear Nancy:

   I use pH8.0 buffer caps made by Metrepak, Brooklyn, NY (Fisher catalog #13640303F).  Carefully pull open capsule, dissolve powder in 100 ml. of distilled water. Add 0 .75 ml of Toluidine blue stock solution (Stock solution is 1% aqueous Tol. Blue...shelf life 6 months).  I stain the deplasticized sections for 18-24 hours, dehydrate in 2 changes each of 95% and 100% ethanol, clear in xylene and coverslip with Permount.  I suppose you could increase the strength of the staining solution and shorten the staining time, but I'm in a research position, and time is usually not an issue!  We've been doing it this way for the last 15 years and had no complaints!!

Hope this helps!
Diane Mahovlic
Othhopedic Pathology Laboratory
The Cleveland Clinic Foundation
Cleveland, Ohio 

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