78 



viscera exposed. (Directions for the post-mortem examina- 

 tion will be given in the course in pathology). 



Inoculate a tube of bouillon, one of agar, and a fermenta- 

 tion tube of glucose bouillon, from either the liver, spleen, 

 or kidney. (In an actual investigation of an unknown dis- 

 ease, cultures should be made from all of the organs, blood , 

 and lymphatic glands). 



Make a series of 3 agar plate cultures. 



Make several cover-glass preparations from the organ from 

 which the cultures were made. 



Stain and examine the cover-glass preparations and describe 

 the bacteria, if any are found. Stain with alkaline methy- 

 lene-blue and carbol fuchsin. (It is sometimes necessary to- 

 preserve pieces of the tissue in alcohol or to fix them in 

 some of the other fixing fluids for sectioning and staining, 

 preparatory to studying them). 



Preserve one or more of the cover-glass preparations to 

 accompany the notes. 



136. Making cultures from animal tissues. Heat a 

 platinum spatula to a red heat in a gas flame and scorch the 

 surface of the organ. Flame a pair of fine forceps and tear 

 the scorched surface away, and with the platinum loop take 

 up a loopful of the tissue underneath with which inoculate 

 the media. It is also desirable to inoculate a tube of slant 

 agar with the needle by drawing it over the surface after 

 changing it with tissue. In making plate cultures use a 

 loopful of the crushed tissue for the first tube. The quantity 

 of the tissue necessary to give a desired number of colonies 

 cannot be anticipated, although experience in working with 

 different organisms in animals renders one able to- 

 approximate the amount required. 



