Tylka Nematology Lab

Procedure:

• Wash the cyst/sediment suspension (from elutriator or manual extraction method) from the 100 ml beakers into the small PVC #60 sieve.

• Place a #200 over #500 sieve into the funnel below the rubber stopper, and start the drill press using the foot pedal.
• Bring the spinning rubber stopper into contact with the sieve surface.

• Grind the material on the sieve lightly between the rubber stopper and sieve. Periodically rinse with the wash bottle. This will maintain lubrication between the sieve and pestle and also rinse eggs through the #200 sieve. Grind until nothing remains upon the small PVC sieve except sand.

• Release pressure on the foot pedal to stop the drill press, wait for the drill press to stop spinning, and rinse the rubber stopper.
• Wash the material on the #500 sieve back into the original 100 ml beaker.

• Carefully rinse all sieves under the faucet.
• Repeat for all remaining samples.
• Place 4 to 5 drops of 1 M HCl and approximately one eye-dropper full of acid fuchsin stain into each beaker of egg solution.
• Heat beakers containing eggs and stain in the microwave until the suspension is about to boil (approximately 3 minutes for 12 samples).
• Remove beakers from microwave and allow to cool.
• Place beakers in the refrigerator until they are counted.

Sources:

• Standard sieves:
• Fisher Scientific
  1600 W. Glenlake Avenue
  Itasca, IL 60143
  (800) 223-9114 or
• VWR Scientific
  P.O. Box 66929, O'Hare AMF
  Chicago, IL 60666
  (800) 932-5000
• The #60 (PVC) sieve:
• David Soh
  (515) 294-5094
• Rubber stopper:
• David Soh
  (515) 294-5094
• Motorized stirrer for rubber stopper:
• Delta 10" drill press running at 2340 RPM (any brand would work).
• Chemicals:
• Fisher Scientific
  1600 W. Glenlake Avenue
  Itasca, IL 60143
  (800) 932-5000