Baker's
Hematal Variants

Ingredient Stock A Stock B Function
Aluminum sulphate 15.76 g Mordant
Distilled water 1 L Solvent
Haematein 1.876 g Dye
Ethylene glycol, 50%, aqu. 1 L Solvent
 
Ingredient Hematal-8 Hematal-16  
Stock A 1 vol 2 vol  
Stock B 1 vol 1 vol  
Ethylene glycol, 50%, aqu. 1 vol  
 
Ingredient Standard A Standard B Function
Aluminum sulphate 3.94 g Mordant
Distilled water 1 L Solvent
Haematoxylin 5 g Dye
Sodium iodate 0.5 g Dye
Distilled water 1 L Solvent
Sulphuric acid, 0.5% aqueous, as needed.
Ammonia, 0.5% aqueous, as needed.

 

Compounding procedure

Hematal-8 and Hematal-16
Prepare each stock solution seperately.
Combine as specified.
The solution may be used immediately.

Standard
Prepare each solution seperately.
Standard solution B should be brought to a boil, then immediately cooled.

 

Method (Hematal-8 & Hematal-16)

  1. Bring sections to water with xylene and ethanol.
  2. Place into the staining solution for the specified time.
    • Hematal-8 for 2-5 minutes.
    • Hematal-16 for 10 minutes.
  3. Rinse with water and blue.
  4. Rinse well with water.
  5. Counterstain if desired.
  6. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

 

Method (Standard)

  1. Bring sections to water with xylene and ethanol.
  2. Place into standard solution A for 1 hour.
  3. Rinse with water.
  4. Place into standard solution B for 20 minutes.
  5. Rinse with water.
  6. Place into 0.5% sulphuric acid for 40 seconds.
  7. Rinse with water.
  8. Place into 0.5% ammonia water for 5 seconds.
  9. Wash well with water.
  10. Optionally, counterstain with eosin.
  11. 70% ethanol for 30 seconds.
  12. 90% ethanol for 30 seconds.
  13. Absolute ethanol, 2 changes, 1 minute each.
  14. Xylene, 2 changes, 1 minute each.
  15. Mount with a resinous medium.

 

Expected results

Notes

  1. Hematal-8 and Hematal-16 may be used for routine staining. The solutions are quickly and conveniently prepared.
  2. Hematal-8 may be used regressively by applying for 30 minutes, then lightly differentiating in acid. Dilute sulphuric acid was recommended (0.5% aqu.), but hydrochloric acid was usable.
  3. Hematal-8 has 8 atoms of aluminum for each molecule of hematein, Hematal-16 has 16, hence the names.
  4. Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.
  5. The standard method was used to give consistent staining with the usually expected appearance, but with the ability to vary each step for experimental reasons. It was not intended for routine use.

 

Reference
Baker, J. R., (1962),
Experiments on the action of mordants: 2. Aluminium-haematein.
Quarterly Journal of Microscopical Science, v. 103, pt. 4, pp. 493-517.

 


 

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