28 PRACTICAL BACTERIOLOGY. 



DEMONSTRATION XI. 



Micrococcus Lanceolatus (Pneumococcus) in Sputum — Bacillus In- 

 fluenzas in Sputum — Sections of Pneumonic Lung — 

 Tissues containing Pyogenic Micrococci. 



Pneumonic Sputum. 



Fresh sputum is furnished when possible, otherwise 

 sputum preserved in 5% carbolic acid solution is used. 

 Films prepared from the preserved sputum have to be 

 cleared, after fixation in the flame, by placing in a 

 mixture of equal parts alcohol and ether for 1 to 2 

 minutes. 



Prepare cover glass films from the sputum by placing 

 a small portion on a cover glass and, after pressing on 

 a second cover glass, sliding the two apart. 



1. Stain the films so prepared, by Gram's method, 

 counterstaining with eosin. 



This method stains the capsule fairly well. 



2. Stain other prepared films of the pneumonic 

 sputum in eosin and methylene blue. 



3. For staining the Pneumococcus and its capsule 

 in fresh sputum, use the following method: 



(a) Prepare films in the usual manner, dry and fix 

 in the flame. 



(b) Pass the films quickly through glacial acetic 

 acid. 



(c) Without washing, place the film in anilin gen- 

 tian violet for 5 to 6 minutes. 



