[Histonet] Toluidine blue staining

[Edwards, Chris]

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

 



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