xv PHAGOCYTOSIS AND GENERAL INFECTION 73 



(c) Wash in distilled water, and dry between filter- 

 paper. 



(d) Pass four to six times through the flame. 



(e) Stain in Loffler's methylene - blue for about ten 

 seconds. 



(/) Wash again, dry and mount. 



Examine with a high power and ^ i* 1 - ^ immersion 

 for phagocytes and eosinophile cells. Observe the 

 relation of the latter to the bacilli. 



(4) Local immunity and general infection. 



(a) Inoculate a pithed frog under the skin of the thigh 

 with a few drops of a fresh broth culture of Bacillus 

 anthracis. 



(b) To do this, with a fine-pointed pair of scissors snip 

 a small hole in the skin of the ventral surface of the thigh, 

 just above the knee. 



(c) With a fine capillary tube blow a small quantity of 

 the broth culture under the skin. 



(d) Half an hour later, open the chest of the frog and 

 expose the heart. 



(e) With a sterile capillary tube remove some of the 

 heart's blood, blow it over the surface of a sloped gelatine 

 tube, and then distribute the blood uniformly by means of 

 a platinum needle. 



(/) Place the tube in the incubator at 22 C. Twenty- 

 four or forty-eight hours later examine it for colonies of 

 Bacillus anthracis and make impression specimens (vide 

 p. 26). 



