Smith's Vanadium Hematoxylin
for Basic Proteins including Histones

When differentiated with dilute lithium carbonate this solution demonstrates basic proteins, including nuclear histones.

Ingredient Amount Function
Hematoxylin 50 mg Dye
Ethanol, 100% 5 mL Solvent
Glycerol 10 mL  
Distilled water 35 mL Solvent
Ammonium meta-vanadate 200 mg Solvent

 

Compounding procedure
Dissolve the hematoxylin in the ethanol, then add the glycerol and water in sequence.
Add the ammonium vanadate and stir for 30 minutes.
The ammonium vanadate may not completely dissolve.
The pH is 6.2

Tissue
Paraffin sections of Schaudinn, Bouin or formalin fixed tissues are suitable. Sections of Zenker or Helly fixed tissues must be soaked in saturated aqueous lithium carbonate for 4 hours to stain satisfactorily.

Method

  1. Bring sections to water with xylene and ethanol.
  2. Place into the vanadate hematoxylin for 30 minutes.
  3. Place directly into 0.08% aqueous lithium carbonate for the appropriate time.
  4. Briefly rinse with distilled water for 2 seconds.
  5. Optionally, counterstain if wished.
  6. Dehydrate with 70%, 95% and absolute ethanols.
  7. Clear with xylene and mount with a resinous medium (Permount specified).

Expected results

Differentiation times in lithium carbonate
Bouin2 minutes
Formalin, 10%2 minutes
Helly10 minutes
Schaudinn5 minutes
Zenker10 minutes

 

Notes

  1. Counterstaining can be accomplished at step 5 with 1% aqueous eosin Y, phloxine B or erythrosin B.
  2. Alternatively, metanil yellow at 0.1% in 95% ethanol containing 0.1% acetic acid may be used between the 70% and 95% ethanols in step 6.
  3. Counterstaining with 0.01% aqueous safranin O stained RNA intensely, giving good contrast between blue nuclear histone and endoplasmic reticulum.

 

Reference
Smith, A. A., (1995)
A vanadate hematoxylin stain for basic proteins.
Biotechnic and Histochemistry, v. 70, Nº 5, p. 5.

 


 

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