RE: Toluidine blue stain for undecalified bone (MMA)

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From:jim <>
To:'Nancy Maronto' <>, "" <>
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Phosphate buffer certainly precipitates with seawater and I guess that the Ca 
in bone reacts with that buffer. I don't know what pH the stain is, but it 
never produced precipitates for me.
I used for years solutions made up from 1% Borax and 0.5% Toloidine Blue; this 
solution keeps for years. A few drops of stain solution is filtered onto a 
resin embedded section and left on a 60 C hotplate until a drying ring forms on 
the stain solution. The add a drop or two of acid alcohol for
differentiation. This is one of the most attractive features of the Toluidine 
stain as it can give a two
colour, pink and blue rendition. Then immediately rinse the slide with 
distilled water. The acid alcohol can destain partially or even completely, but 
a brief application brings forth the two colours.

Make acid alcohol by adding to 25ml of 50% Ethanol a couple of drops of 1N HCl.
Jim Darley

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On Friday, August 25, 2000 4:12 AM, Nancy Maronto [] 
> I am looking for a procedure for T blue stain at PH of 8 for undecalcified
> bone in MMA.  These are thin sections (5 microns).  I tried the procedure in
> Sheehan's Histotechnology book but used a phospate buffer to reach the pH.
> This caused a problem with what I think was precipitate on the slide.  Any
> suggestions?
> Thanks,
> Nancy Maronto
> MPI Research
> Mattawan, MI
> (616)668-3336

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