STAINING, PRACTICAL AND THEORETICAL 



MALLORY'S PHOSPHOTUNGSTIC ACID HAEMATOXYLIN 

 A general stain for vertebrate tissues 



Solution required: 



Haematoxylin io% in absolute 

 alcohol (ripened for three 

 months or longer) . . . . i ml. 



Phosphotungstic acid . . . . 2 gm. 



Distilled water . . . . . . 100 ml. 



Note: If ripened haematoxylin solution is not 

 available, the following artificially ripened stain 

 should be used: 



Haematoxylin (dry) o-i gm., phosphotungstic acid 

 2 gm., distilled water 100 ml., potassium perman- 

 ganate 1% aqueous 1-77 ml. 



Technique: 



1. Fix in Zenker. Embed in paraffin wax. 



2. Bring sections down to distilled water. 



3. Treat with iodine to remove mercuric precipitate. 



4. Remove iodine with 0-5% aqueous sodium hyposulphite. 



5. Wash thoroughly in running water. 



6. Immerse for five to ten minutes in 0-25% potass, perman- 

 ganate ; then wash in tap water. 



7. Immerse for ten to twenty minutes in 5% oxalic acid; then 

 wash thoroughly with tap water. 



8. Stain twelve to twenty-four hours in haematoxylin solution, 

 prepared as above. 



9. Wash in tap water; dehydrate with 95% and absolute 

 alcohol. 



10. Clear in xylol and mount. 



Results: 



Nuclei, centrioles, achromatic spindles, fibroglia, myoglia, 

 neuroglia fibrils, fibrin, contractile elements of striated muscle: 

 blue. Collagen, reticulum, ground substances of cartilage and bone: 

 yellowish to brownish red. Coarse elastic fibrils : faint purple. 



References : 



Jones, Ruth McClung (1950), p. 244. 

 Mallory, F. B. (1900). 



288 



