A Technique to Inflict Closed-Head Traumatic Brain Injury in Drosophila Flies

Take control and test adult Drosophila flies under anesthesia.

Place the test fly in a holder and gently tap it to position the fly's head outside the tip. Ensure the fly's mouthparts and body remain inside the holder to avoid non-target injuries.

Attach the holder to a strike device with the fly's head facing downward.

The device is connected to a controlled gas supply and contains a gas-propelled impactor to inflict injury.

Deliver a gas burst to propel the impactor and strike the fly's head, inducing a closed-head traumatic brain injury without external damage.

The resulting mechanical stress damages neurons, impairing their function.

Detach the holder to release the fly, then transfer it to a vial for recovery.

For the control fly, apply a gas burst without striking it with the impactor.

The control and test flies are now ready to assess the impact of the injury.

Anesthetize a single two-day-old adult female fly using carbon dioxide.

From the carbon dioxide pad, use a fine brush to gently transfer the fly into the holder.

Then, gently tap the holder until the head protrudes from the tip.

If the proboscis is exposed, use a blunted 1 milliliter syringe needle to gently tuck it ba