164 
Dourine 
of iodized glycerine to 9-0 c.c. serum. At the same time one should 
collect and store a number of negative control sera under the same 
conditions. 
II. Titration of Reagents. 
(1) Titration of Haemolytic Serum ( Amboceptor). 
Prepare the following stock dilutions of serum and corpuscle 
suspension: 
1. Haemolytic serum (rabbit anti-sheep) . . 0-1 cc. 
Normal salt solution (0-85 per cent.) . . 9-9 „ 
10 0 „ .... 1 : 100 
2. Complement: 
Fresh guinea-pig’s serum .... 1-0 ,, 
Normal salt solution ..... 19-0 „ 
20 0 „ .... 1:20 
3. Corpuscle suspension: 
Washed sheep’s corpuscles (50 per cent, stock 
suspension) ..... 2-0 „ 
Salt solution ...... 23-0 ,, 
250 „ ....1:25 
Further dilutions of the haemolytic serum are made as under: 
Salt Haemolytic 
solution serum 
c.c. c.c. 
Tube No. 1 
3-0 
1-0(1: 100) e 
quals 1 : 400 (0-0025 serum in 
1-0 c.c.) 
2 
11 " 
5-0 
i-o 
11 
1 : 600 (0-0016 
V 
„ ) 
„ 3 
7-0 
10 „ 
11 
1 : 800 (0-0012 
11 
„ ) 
„ 4 
9-0 
10 
11 
1 : 1000 (0-001 
IT 
„ ) 
„ 5 
0-5 
1-0 (1 : 1000) 
11 
1 : 1500 (0-00066 
11 
„ ) 
„ 6 
10 
1-0 „ 
11 
1 : 2000 (0-0005 
11 
„ ) 
„ 7 
20 
1-0 
19 
1 : 3000 (0-00033 
1) 
„ ) 
„ 8 
3-0 
i-o 
1) 
1 : 4000 (0-00025 
91 
» ) 
„ 9 
4-0 
i-o 
19 
1 : 5000 (0-0002 
„ ) 
In each tube 1-0 c.c. only of the dilution is held back, the excess 
amount being discarded, 1-0 c.c. each of complement and red cell sus¬ 
pension added, which with 2-0 c.c. salt solution make a total volume 
of 5-0 c.c. in each tube. 
The complete titration set is then: 
Serum 
dilutions 
Salt 
Complement 
Red Cell 
(as above) 
solution 
1 :20 
suspension 
c.c. 
C.C. 
c.c. 
c.c. 
Tubes 1-9 
10 
2-0 
10 
1-0 
Titration set 
Tube No. 10 
1-0 
3-0 
— 
1-0 
Serum control (1 : 100 dil.) 
„ 11 
— 
3-0 
10 
1-0 
Complement control 
12 
— 
4-0 
— 
1-0 
Red cell control 
