THE PRODUCTION OF A HEMOLYTIC SERUM 323 



quantity of salt solution so that the dilution may be approx- 

 imated. 



6. Give results and conclusions in full. 



REFERENCES 



MCFARLAND: Pathogenic Bacteria and Protozoa, 7th Ed., p. 146. 



MARSHALL: Microbiology (1911), pp. 570-574. 



NUTTALL: Blood Immunity and Relationship (1904). 



KOLMER: Infection, Immunity and Specific Therepy (1915), pp. 70, 71, 



292-315, 517-519, 844-846. 

 ZINSSER: Infection and Resistance (1914), pp. 248-271, 



EXERCISE 13. THE PRODUCTION OF A HEMOLYTIC 



SERUM 



For this work a rabbit will be immunized to washed 

 sheep blood cells. 



Apparatus. Sterile physiological salt solution; glass 

 beads; small glass funnel; two 200 c.c. Erlenmeyer flasks; 

 five or six sterile centrifuge tubes; sterile 5 c.c. pipette, 

 with rubber bulb attached for draining off serum and salt 

 solution in centrifuge tubes; sterile 14-gage 2j-inch hypo- 

 dermic needle ; sheep; rabbit. 



Note. Chemically pure salt and distilled water should be used in 

 the preparation of salt solution for this work and for the complement 

 fixation test. 



Method. 1. In one Erlenmeyer flask place eight or 

 ten glass beads for defibrinating blood, plug and sterilize 

 in hot air. 



Provide the other one with a small, sterile, glass funnel 

 and two layers of sterile cheese-cloth for filtering and de- 

 fibrinating blood. 



2. With an attendant holding the sheep, clip the wool 

 over the area of the jugular vein and wash with 50% alcohol. 



3. Place the thumb over the jugular furrow about half 

 way between the head and shoulder and press on the vein 

 so that it will become distended. 



