Fly Aspirator Tube: A Simple Tool to Transfer Live Drosophila

протокол

This protocol is an excerpt from Barradale et al., Quantification of Drosophila Grooming Behavior, J. Vis. Exp. (2017).

1. Prepare an aspirator for moving live Drosophila from a culture vial to the grooming chamber. Aspirators allow transfer of conscious animals to the behavioral chambers to ensure that anesthesia does not affect subsequent behavioral observation.
   1. Using scissors cut 1.5 feet of tygon tubing ID 1/8", OD 1/4". Cut at least 1 inch off of the tip of a 1 mL disposable micropipette tip. Hold a 1 cm square piece of mesh (opening 0.196 inch) over the cut tip.
      NOTE: Most 1mL tips have a gradation line where the tip sits in the package box, typically cut to that line.
   2. Snugly place a fresh 1 mL micropipette tip over the mesh/cut tip. Observe the mesh form a tight barrier between the two tips. The inside of the new tip creates a holding chamber for flies.
   3. Cut the extreme tip of the new micropipette tip to widen the opening enough to allow passage of a single fly. The hole will roughly match the opening of the grooming chamber (approximately 1.5-2 mm in diameter).
   4. Snugly fit the nested tips onto the end of the tygon tubing. Push the tips into the tube to ensure a tight fit to allow for vacuum pressure.
   5. On the other end of the tubing, cut the tip of a 200 μL micropipette tip and fit the narrow cut end into the tubing. This is the "mouth" side of the aspirator.

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Материалы

Name	Company	Catalog Number	Comments
High-Flex Tygon PVC Clear Tubing	McMaster-Carr	5229K54	ID 1/8", OD 1/4", used with micropipettor tips and mesh to construct mouth aspirators
Micropipette tips (1 mL and 200 μL)	Genesee Scientific	24-165, 24-150R
Nylon Mesh Screen, 2 x 2.6"	McMaster-Carr	9318T44	Used to construct grooming chamber and mouth aspirators