86 



LABORATORY COURSE IN SERUM STUDY 



The following test is made to determine the haemolytic power of the 

 anti-cat rabbit serum : 



In testing the agglutinative power set up a similar series but without 

 fresh serum. 



II. IN Vivo EFFECT OF H^MOLYTIC SERUM 



Etherize two cats and inject 1 c.c. of immune serum into one and 

 3 c.c. into the other. Give the injections directly into the saphenous 

 vein with a fine hypodermic needle. Put the first cat into a clean cage 

 with a dram for the collection of urine after 6, 24 and 48 hours. (The 

 first urine will con tain large amounts of haemoglobin, the last bile.) Half 

 an hour after the injection collect a specimen of the cat's serum by prick- 

 ing the ear vein. Centrifugalize and observe the blood-tinged serum. 

 Make smears of the blood, stain, and note with oil immersion lens the 

 phagocyted red cells in some of the leucocytes. The second cat will 

 probably die at once. If so, autopsy immediately and look for clumps 

 of agglutinated red cells in portal and pulmonary veins. 



To demonstrate the in vivo effect of complement, give a rabbit an 

 intravenous injection of 2.0 c.c. of 25 % suspension of sensitized sheep 

 blood cells. (In sensitizing the cells, regard 0.1 c.c. of 25 % cells as a 

 unit and add two units of amboceptor for every unit of cells. After 

 the cells have stood in contact with the amboceptor for one half hour, 

 centrifugalize them and make them up to the original volume of 25 % 

 suspension). 



After a half hour collect a specimen of serum from the rabbits' 

 ear and examine for dissolved haemoglobin. Make a smear of the blood 

 and examine for phagocytosis. Examine the rabbit's urine after 6, 

 24 and 48 hours. 



The experiments in this lesson need not be done by the entire class 

 but can be done as a demonstration by a small group of students working 

 with the instructor. 



