RE: Toluidine blue stain for undecalified bone (MMA)
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From: | jim <jim@proscitech.com.au> |
To: | 'Nancy Maronto' <nmaronto@hotmail.com>, "histonet@pathology.swmed.edu" <histonet@pathology.swmed.edu> |
Reply-To: | |
Content-Type: | text/plain; charset=us-ascii |
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.
Cheers
Jim Darley
ProSciTech Microscopy PLUS
PO Box 111, Thuringowa QLD 4817 Australia
Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service@proscitech.com
Great microscopy catalogue, 500 Links, MSDS, User Notes
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On Friday, August 25, 2000 4:12 AM, Nancy Maronto [SMTP:nmaronto@hotmail.com]
wrote:
> 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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