118 LABORATORY COURSE IN SERUM STUDY 



to whether the alcoholic extract is run rapidly into the salt solu- 

 tion which is used as diluent or whether the extract is diluted by 

 dropping salt solution into it. It is not of so great importance 

 which of the two methods is used as that the same method should 

 be used throughout both preliminary titrations and tests. 



Each student tests one of the three antigens made by the class. 

 This antigen is to be made up to a 1-10 dilution with saline. Add the 

 saline slowly to the antigen. Each student needs about 10 c.c. of the 

 1-10 dilution. 



A syphilitic (Wassermann-f + + +) and a normal human serum con- 

 taining no or very little antisheep-cell amboceptor are chosen : they are 

 inactivated at 56 C. for half an hour before use. 



Set up the following series of tubes : 



1. DETERMINATION OF ANTICOMPLEMENTARY DOSE 



Tube Antigen 1-10 Complement 1-10 



1 0.2 c.c. 0.5 c.c. 



2 0.4 c.c. 0.5 c.c. 



3 0.8 c.c. 0.5 c.c. 



4 1.2 c.c. 0.5 c.c. 



5 1.6 c.c. 0.5 c.c. 



6 2.0 c.c. 0.5 c.c. 



2. DEMONSTRATION THAT ANTIGEN is NOT HJEMOLYTIC OF ITSELF 

 Tube 7 Antigen 1-10 1.0 c.c. 5 % Sheep cells 0.5 c.c. 



3. DETERMINATION OF ANTIGENIC DOSE 



Tube Antigen Syphilitic (+ + + +) Serum Complement 1-10 



8 (1-40) 0.05 c.c. 0.1 c.c. 0.5 c.c. 



9 (1-40) 0.1 c.c. 0.1 c.c. 0.5 c.c. 



10 (1^0) 0.2 c.c. 0.1 c.c. 0.5 c.c. 



11 (1-10) 0.1 c.c. 0.1 c.c. 0.5 c.c. 



12 (1-10) 0.2 c.c. 0.1 c.c. 0.5 c.c. 



13 (1-10) 0..4 c.c. 0.1 c.c. 0.5 c.c. 



4. PROOF THAT ANTIGEN is SPECIFIC (NEGATIVE CONTROL) 



Tube Antigen 1-10 Normal Human Serum Complement 1-10 



14 0.1 c.c. 0.1 c.c. 0.5 c.c. 



15 0.2 c.c. 0.1 c.c. 0.5 c.c. 



16 0.4 c.c. 0.1 c.c. 0.5 c.c. 



