Jaclynn,
I haven't tried this with T Blue, but Azure B (another metachromic stain)
requires different ph's to stain different tissue components.  I use a 0.2%
solution of Azure B and make three different ph solutions.  AZ B at ph1.2 (acid
ph's) stains some matrix tissue like cartilage selectively, AZ B at ph 4 will
stain cartilage and bone components and AZ B at ph 7 stains just about
everything.  My point is, you may have to use different ph solutions for
different tissue compentents.  I use dilute sodium hydroxide or acidic acid to
ph my solutions.
Patsy Ruegg

Jaclynn Lett wrote:

> Regarding my Toluidine Blue stain solution, which should be staining animal
> tissue varying shades of blue-purple (which I want), but is now staining
> bright blue instead (which I don't want):
>
> 1% Toluidine Blue O
> 1% Sodium Tetraborate Dodecahydrate
> 30% Ethanol
> in DDI
>
> (1g. Tol.Blue, 1g Borax, 30mL Ethanol in 100mL DDI)
>
> I've received several suggestions to change the pH--some to acidic, some to
> basic.  Which direction should it be changed and what chemical do I use to
> change it?
>
> Thank you again,
>
> 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
>
> jlett@cid.wustl.edu
> 314-977-0257