13 



the following information regarding the technique of the 

 best methods of staining the organism are given, together 

 with the composition of the reagents required : — 



(i) Nicolle's Violet — 



Saturated solution of gentian violet in 90 per cent. 



alcohol loc.c. 



I per cent, aqueous solution carbolic acid . .100 c.c. 



(2) Nicolle's Thionine — 



Saturated solution of thionine in 90 per cent, alcohol 10 c.c. 

 1 per cent, aqueous solution of carbolic acid . lOO c.c. 



(3) Solution for Gram-Weigert-Kuhne's method — 



Concentrated solution of crystal violet . . . i part. 



Distilled water, to which a few drops of hydrochloric 



acid have been added 10 parts. 



(4) Gram's Iodine solution (Lugol's solution) — 



Iodine i part. 



Iodide of potassium 2 parts. 



Distilled water 3°° P''»rts. 



(5) Kuhne's Iodine solution — 



Iodine 2 parts. 



Iodide of potassium ...... 4 P^rts. 



Distilled water 100 parts. 



(6) Counter stain — 



Saturated solution of vesuvine (Bismarck brown). 



(7) Decolourizing agents — 



Alcohol or aniline oil. 



(8) Clarifying agents — 



Zylol or clove oil. 



(9) Mounting agents — 



Canada balsam or Farrant's solution. 



Gram-Nicolk method. 



Make a thin smear of pus from the suspected 

 wound, ulcer or pustule, on a cover-glass or slide, fix it 

 in the ordinary manner by passing it three times through 



