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- A four-way olfactometer is a closed chamber with one-way ports at each corner, through which it can deliver up to four odors, creating quadrants with distinct odor profiles. Begin an experiment by transferring awake flies into the chamber. Avoid anesthesia, as this may affect their behavior during the experiment.
Expose the flies to clean air and observe their movements to make sure that no odorants are contaminating the olfactometer and affecting fly behavior. Then, prepare the test odorants in separate vials and connect them to the main chamber through the one-way ports. Expose the flies to the odorants for a pre-defined period, removing the stimulus after the trial has ended.
Flies tend to avoid quadrants with repellent odors while moving towards and settling in quadrants with attractive odors. Measure the flies' odor-cued behavioral responses by tracking their positions over time as they explore the olfactometer.
In the following protocol, we will use a four-quadrant olfactometer with an air delivery system to analyze fly behavior in response to ethyl propionate, a repellent odorant, and apple cider vinegar, which is attractive.
- At least five minutes in advance, switch on t
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