Section III: Examine of Phlebotomine Sand Flies for Leishmania
Dissecting the Gut
- Transfer flies to the tube of soap solution and shake gently;
- Pour contents of tube onto a fine-mesh screen stretched over the mouth of a large beaker and rinse with tap water until soap is gone;
- Using a dissecting pin, lift the flies from the mesh screen and place them in a small Petri dish containing 1x PBS to rinse;
- With the 1cc syringe, apply a drop of PBS onto the glass microscope slide;
- Place a freshly washed/rinsed sand fly in the drop of PBS on the microscope slide;
6. With two dissecting needles (straight one in the left hand and bent one in the right hand), remove the sand fly's legs;
7. Steady the fly by piercing the thorax at the base of the wing with the straight pin and holding it against the glass; with the bent pin in the right hand, remove the sand fly's head;
8. Keeping the pin in the left hand in position to secure the fly, press the tip of the bent pin horizontally against the tip of the abdomen between segments 7 and 8 and pull firmly but slowly and steadily downward (toward you) until the gut comes out of the abdomen.
9. If the end of the gut breaks off, place the bent segment of the pin horizontally across the anterior abdomen and squeeze the gut out of the abdomen, as you would toothpaste out of a tube.