STAINING, PRACTICAL AND THEORETICAL 



is allowed to stand for longer than thirty minutes, a precipitate is 

 liable to form around the leucocytes, making accurate counting 

 impossible. 



For more detailed information, the original paper should be 

 consulted. 



Reference: Moore, J. A. Ill & James, G. W. Ill (i953). 



TRIBASIC STAIN (Barlow) 

 For mast- cell granules 



Solutions required: 



A. Iron alum, 5% aqueous . . . . 100 ml. 

 Celestin blue 0-5 gm. 



Shake the iron alum solution with the dye in a 

 flask; boil for three minutes, allow to cool; then 

 filter, and add: 



Sulphuric acid, cone. . . . . 2 ml. 



Glycerine . . . . . . . . 14 ml. 



B. Haemalum (Mayer) 



C. Lithium carbonate, saturated, 

 aqueous 



Alcohol, 70% 99 ml. 



HCl, cone. . . . . . . I ml. 



Acridine red, 10% aqueous 

 Neutral red, 2% aqueous 

 Basic fuchsin, 1% aqueous 

 Barlow's tribasic stain 

 Solution E 

 Solution F 



D 



E. 

 F. 

 G 

 H 



Solution G 

 Distilled water 

 Nitric acid, cone. 



10 ml. 



25 ml. 



5 ml. 



20 ml. 



0-25 ml. 



Boil the mixture vigorously for five to ten minutes ; 

 cool; then add: 

 Glycerine . . . . . . . . 50 ml. 



Wash the glycerine out of the measuring cylinder 

 with about 25 ml. of distilled water; add the washings 



448 



