Re: Toluidine Blue problem

From:Teresa Flores

Jaclynn, The Journal of Histotechnology/vol.21,No.3/September 1998 Pg 213.
Please read Staining (pg 214) where a 2-step stain, Toluidine Blue "O"
Stain and a Basic Fuchsin Stain is used on Spurr epoxy sections. Our EM lab
has used this stain since Paragon was removed from the shelves. I do agree
that there needs to be an "ripening time" for the stain. I have always
prepared batches of 2,500 ml and filtered from there as needed. Please give
this procedure a try. Receipe DOES NOT contain any ethanol. Let me know how
your pathologist likes it.

>I don't know how many of use cut plastic sections, but I have a question
>regarding a Toluidine Blue stain solution:
>1% Toluidine Blue O
>1% Sodium Tetraborate Dodecahydrate
>30% Ethanol
>in DDI
>I've been using this stain for years on Epon-Araldite sections and Durcupan
>sections with no problems in getting satisfactory staining.  When I have
>30-50mL remaining, I always make a new batch so it's ready the next time I
>need it , but this time I forgot.  I made a new solution and had to use it
>right away.  I stained my sections as usual, but the color of the sections
>was bright blue instead of the more purple-blue we desire.  Thinking that
>I'd made a mistake in preparing the solution, I made another batch and
>decided to wait overnight to try it, but again the sections were bright
>I was never aware of the need to do this, but does this solution need to
>"ripen" for a week or so before using?  I can't think of any other reason.
>The quality of our water hasn't changed, and everything else is the same as
>I've always used.
>Thank you for any help you can give me.
>Jaclynn M. Lett, Research Technician
>Electron Microscopy Core Facility
>Fay and Carl Simons Center for Biology of Hearing and Deafness
>Central Institute for the Deaf
>4560 Clayton Ave.
>St. Louis, MO 63110

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