DIRECTIONS FOR THE USE OF STAINS 13! 



5. Loffler's Flagella Stain: 



(a) Preparation of the Mordant. Dissolve 2 grams of 

 desiccated tarmic acid in 15 c.c. of distilled water 

 (heat gently), and add 5 c.c. of a saturated solution 

 of ferrous sulphate, i c.c. of an alcoholic solution 

 of fuchsin (i gram of fuchsin, 25 c.c. warm ab- 

 solute alcohol), i c.c. of i per cent, sodium hydrate 

 solution. Filter. 



(a) Preparation of Stain. One part of a saturated alco- 

 holic solution of fuchsin to 4^ parts of a 5 per cent, 

 solution of carbolic acid. 



0) Filter. 



(a) Method of Staining. Mordant one minute. 



(b) Wash in distilled water. (Have water in wide- 



mouthed beaker.) 



(c) Stain with carbol-fuchsin one-half minute (heat 



gently). 



(d) Wash in water. 



(e) Dry thoroughly. 



(f) Treat with xylol. 



(g) Mount in balsam. 



6. Zettnow's Flagella Stain: 



(a) Solution (i): Dissolve 2 grams of tartar emetic 



(2K(SbO)C 4 H 4 O 6 + H 2 O) in 40 c.c. of water. 



(b) Solution (2) : Dissolve 10 grams of tannic acid 



(Ci 4 HioO 9 ) in 200 c.c. of water. 



(c) Warm solution (2) to 50 to 60 C., add 30 c.c. of the 



tartar emetic solution (i). The turbidity of the 

 mordant should entirely clear up on heating. 



(d) Next dissolve i gram silver sulphate in 250 c.c. 



distilled water. 



