Groups of 50 male flies were collected 0-1 days after eclosion and trained at 3-5 days of age. Flies were trained in vials (2.5 cm diameter and 9.5 cm height) containing 1% agar. Vials contained 64 evenly-spaced perforations and a mesh lid to allow even distribution of ethanol. Vials were placed into a holder in a 30 cm length × 15 cm height × 15 cm width training chamber. The training chamber (Aladin Enterprises, Inc., San Francisco, CA) had three nozzles to allow for air/odorants/ethanol to stream in and one nozzle for waste to stream out.
