TISSUE MEDIUM 215 



men with all aseptic precautions, remove kidneys and testicles and 

 transfer to a sterile glass dish. Cut up the organs with sterile 

 scissors into small pieces say 4 millimetre cubes. The four organs 

 should yield from 25 to 30 pieces of tissue. 



3. Drop a small piece of sterile tissue into the bottom of each 

 sterilised tube. 



4. Take a flask containing about 400 c.c. nutrient agar (+ 10 

 reaction) , liquefy the medium by heat and cool in a water bath 

 to 50 C. 



5. Add 200 c.c. ascitic or hydrocele fluid (horse or sheep serum 

 may be employed, but' is not so good) to the liquid agar and mix 

 carefully to avoid formation of air bubbles. 



6. Fill about 20 c.c. of the ascitic agar into each of the sterilised 

 tubes which already contains a piece of sterile rabbit's tissue, stand 

 all the tubes upright in racks or a j ar, and allow agar to set. 



7. After solidification pour sterile paraffin oil on the surface of 

 the medium in each tube to the depth of 3 centimetres. 



8. Incubate tubes at 37 C. for several days and discard any 

 which prove to be contaminated. 



9. Store such tubes as are sterile for future use. 



