24 LABOKATORY COURSE IN SERUM STUDY 



1. Dog serum unheated (1-10) 0.5 c.c. + Guinea pig cells 2 % 0.5 c.c. 



2. Dog serum unheated (1-10) 0.5 c.c. + Human cells 2 % 0.5 c.c. 



3. Dog serum heated (1-10) 0.5 c.c. + Guinea pig cells 2 % 0.5 c.c. 



4. Dog serum heated (1-10) 0.5 c.c. + Human cells 2 % 0.5 c.c. 



5. Salt solution 0.5 c.c. + Guinea pig cells 2 % 0.5 c.c. 



6. Salt solution 0.5 c.c. -f Human cells 2 % 0.5 c.c. 



Incubate the tubes one hour at 37 C. in water bath, observing 

 changes in cells at intervals of 5, 10, 15, 30, and 60 minutes. Tabulate 

 the results. Tube 1 should show prompt haemolysis; tube 3 should 

 show agglutination of the cells. In tubes 2 and 4 there should be no 

 observable change in the cells in most samples of dog serum. 



Each student needs: 10 J^-inch test tubes, test tube rack, 5 one-c.c. 

 pipettes, 1 five-c.c. pipette. 



LESSON III 



HAEMOLYSIS. (EHRLICH AND MORGENROTH EXPERI- 

 MENTS) 



1. To a group of students is assigned the task of bleeding a sheep 

 from the jugular vein into a flask containing glass beads. After defibrina- 

 tion the cells are washed three times in salt solution and a 5 per cent 

 suspension of the well-packed sediment is made (1.0 c.c. of sediment and 

 19.0 c.c. of salt solution). 



2. To another group of students is assigned the bleeding of guinea 

 pigs from the carotid artery into centrifuge tubes. The blood is allowed 

 to clot, this clot is "rimmed" and the serum separated. The process 

 may be hurried by centrifugation. The serum is then diluted, 1 part of 

 serum to 9 parts of salt solution. 



3. Immune rabbit serum (lytic for sheep cells) will be given out 

 diluted. 1 This serum has been heated for one half hour at 56 C. 

 before dilution, in order to destroy its alexin or complement. This is 

 done so that no unknown amount of complement may be present in the 

 final tests. 



All these materials must be prepared before the regular class period, 

 as the experiments themselves consume several hours. Provided the 

 guinea pig serum is kept very cold (0-^i C.), all the materials can be 

 prepared the preceding day. 



1 In order to save time in the course an immune serum previously prepared by 

 the instructor is used here. The immune haemolytic serum whose preparation 

 was begun by the students in Lesson I is to be titrated later and used for the 

 lessons on complement fixation . 



