FLAGELLA STAINS 103 



9. Wash well in water. 



10. Dry and mount. 



2. "Pitfield" Original Method. 



(a) Mordant. 



Tannic acid i gramme 



Water 10 c.c. 



(b) Stain. 



Saturated aqueous solution of alum 10 c.c. 



Saturated alcoholic solution of gentian violet. . i c.c. 

 Distilled water 5 c.c. 



Mix equal parts of a and b before using. 



1. Prepare and fix the film in the manner described above. 



2. Boil the mixture and immerse the cover-slip in it, whilst 

 still hot, for one minute. 



3. Wash in water. 



4. Examine in water; if satisfactory, dry and mount in Canada 

 balsam. 



3. MacCrorrie's Method. 



Mordant-Stain. 

 Measure out and mix. 



Night blue, saturated alcoholic solution . . 10 c.c. 

 Potash alum, saturated aqueous solution . 10 c.c. 

 Tannin, 10 per cent, aqueous solution . . 10 c.c. 



NOTE. The addition of gallic acid, o.i to 0.2 gramme, may 

 improve the solution, but is not necessary. 



METHOD. 



1. Prepare and fix the films as above. 



2. Pour some of the mordant-stain on the film and warm 

 gently, high above the flame, for two minutes (or place in the 

 "hot" incubator for a like period). 



3. Wash thoroughly in water. 



4. Dry and mount. 



4. Loeffler's Method. 



(a) Mordant. 



Tannic acid, 20 per cent, aqueous solution . . 10 c.c. 



Ferrous sulphate, saturated aqueous solution . 5 c.c. 



Haematoxylin solution 3C.c. 



Carbolic acid, i per cent, aqueous solution . . 4 c.c. 



This solution must be freshly prepared. 



Hcematoxylin solution is prepared by boiling i gramme logwood 



