[Histonet] Toluidine blue staining

Edwards, Chris Chris.Edwards <@t> nationwidechildrens.org
Fri Jun 5 13:15:58 CDT 2009

Hello. Our lab has had highly variable results using toluidine blue to
stain 1.5 micron thick cross sections of plastic embedded nerve tissue
from both mouse and human nerve biopsy. It seems like the stain is
darker in the biopsy tissue than the mouse but both sometimes stain
equally poorly.  We have tried changing the protocol recipe to make it
darker by adding more toluidine blue powder but that has not worked
either. We have used 2 grams toluidine blue, and 2 grams of sodium borax
with 100 ml of distilled water. We have tried changing the sodium borax.
Sodium tetraborate-decahydrate and a sodium tertaborate 99.9% metals
basis from sigma Aldrich have both had the same results. Does anyone
have any idea how to make it stain darker? I can't really emphasize
enough how light the blue is. I did read one protocol that involved
adjusting pH, is that a possible factor? Thanks












Christopher J. Edwards

Research Associate, Sahenk Lab

Center for Gene Therapy

Wexner Building 2, Room 3224

Lab Phone: 355-5224


----------------------------------------- Confidentiality Notice:
The following mail message, including any attachments, is for the
sole use of the intended recipient(s) and may contain confidential
and privileged information. The recipient is responsible to
maintain the confidentiality of this information and to use the
information only for authorized purposes. If you are not the
intended recipient (or authorized to receive information for the
intended recipient), you are hereby notified that any review, use,
disclosure, distribution, copying, printing, or action taken in
reliance on the contents of this e-mail is strictly prohibited. If
you have received this communication in error, please notify us
immediately by reply e-mail and destroy all copies of the original
message. Thank you.

More information about the Histonet mailing list