158 BACTERIOLOGY. 



Method No. XII. Staining of Spores. 



(a) Make a cover-glass spread, dry and pass three times through the 

 flame. 



(b) Add aniline-water gentian-violet. 



(c) Heat until the preparation begins to boil; remove for a minute. 

 Repeat this process six times. 



(d) Wash in three per cent hydrochloric acid-alcohol one minute 



(e) Wash in water. 



(f) Counter-stain with aqueous methylene blue half a minute. 



(g) Wash in water. 



(h) Dry and clear up with xylol. 

 (i) Mount in balsam. 



Method No. XIII. Staining of Flagella. 



(a) Mix upon the cover-glass a portion of the culture with a drop of 

 water, using care not to break off the delicate flagella. 



(b) Dry and pass three times through a flame. 



(c) Apply Loffler's mordant one minute, warming gently. 



(d) Wash in water. 



(e) Stain with aniline-water fuchsin. 



(f) Wash in water. 



(g) Dry and mount in balsam. 



Method No. XIV. Grain's Method for Bacteria. 



(a) Make a cover-glass preparation by the usual method. 



(b) Stain with aniline-water gentian-violet solution, two to five min- 

 utes, warming slightly. 



(c) Add Gram's iodine solution one and one-half minutes. 



(d) Apply alcohol, repeatedly, as long as stain continues to come 

 away from the preparation. 



(e) Wash in water and examine as a water-mount. 



(f ) If desired, dry and mount in balsam. 



Method No. XV. Gabbet's Method for Tuberculosis, etc. 



(a) Make a cover-glass smear of the sputum, pus, blood, or urine to 

 be examined. After the preparation is dry, affix by passing three times 

 through the flame. 



(b) Using a Cornet forceps, apply carbol-fuchsin five to ten minutes, 

 heating until steam appears. 



(c) Wash in water. 



(d) Apply alkaline methylene blue for one minute. 



(e) Wash in water. 



(f) Dry and mount in balsam. 



