STAINING, PRACTICAL AND THEORETICAL 



5. Stain in solution B (toluidine blue) for one hour, in a closed 

 staining dish to prevent evaporation of alcohol. 



Note: Solution B should be filtered before use. 



6. Rinse briefly in solution C to remove excess dye, until the 

 sections appear colourless to the naked eye. 



7. Counterstain lightly by immersing in solution D (eosin) for 

 a few seconds. 



8. Dehydrate rapidly in absolute alcohol. 



9. Clear in xylol and mount in D.P.X., Cristalite, Clearmount, 

 or Emexel, etc. 



Results : 



Mast cells are selectively stained deep blue against a pale pink 

 background. 



Reference: Smith, E. W. & Atkinson, W. B. (1956). 



TOLUIDINE BLUE - SAPONIN METHOD 



For enumeration of basophil leucocytes 



(After J. E. Moore III & G. W. James III, 1953) 



Solutiotis required: 



A. Sodium chloride, A. R 8-5 gm. 



Distilled water 950 ml. 



Dissolve, then make up the volume to i litre with 

 distilled water. 



B. Sodium hydroxide N/io 



C. Solution A 200 ml. 



Test and adjust, if necessary, to pH 7-0 by adding 

 about o-i6 ml. of N/io NaOH, or simply by shaking 

 in two Michrome buffer tablets, pH 7-0, which 

 should first be crushed to powder. 



Toluidine blue 0-15 gm. 



Alcohol, 95% 55 ml- 



446 



