Roach & Smith's Bismuth Hematoxylin

This method demonstrates arginine.

Ingredient Amount Function
Hematoxylin 50 mg Dye
Ethanol, abs. 5 mL Solvent
Sodium iodate 10 mg Oxidant
Bismuth nitrate 50 mg Mordant
Citric acid, 0.5M aqu. 85 mL Acidifier
Glycerol 20 mL Solvent
Sodium hydroxide, 1 M aqu. 19 mL Alkaliser


Compounding procedure
Dissolve the bismuth nitrate into the citric acid solution.
Add 10 mL of the glycerol, then the sodium hydroxide solution.
Adjust the pH to 5.2.
Dissolve the hematoxylin into the ethanol.
Combine the two solutions.
Add the sodium iodate dissolved in 1 mL distilled water.
Stir for 20 minutes.
Add the remaining 10 mL of glycerol.
This solution has a usable life of about 3 hours.


8µ paraffin sections of Bouin fixed tissues are suitable.



  1. Bring sections to water with xylene and ethanol.
  2. Place into bismuth hematoxylin for 10 minutes.
  3. Rinse with water.
  4. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.


Expected results



  1. The formula calls for bismuth nitrate pentahydrate, Bi(NO3)3.5H20.
  2. This method demonstrates proteins with an arginine content of about 12% or more.
  3. Nuclear histones and myelin basic protein stain strongly.
  4. The authors suggest that the bismuth binds to the guanidino groups of arginine.


Roach, J. B., and Allen, A. A., (1997)
Bismuth hematoxylin stain for arginine residues.
Biotechnic & Histochemistry, v.72, Nº 1, p. 49.




Translate in
Google Translate