A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

Take anesthetized adult Drosophila flies in an empty vial.

Insert a cotton ball into the vial to confine the flies at the bottom, ensuring a stronger impact when inflicting injury.

Once the flies recover from anesthesia, attach the vial to the spring of the high-impact trauma device.

Rest the end of the vial on a raised platform. Then, pull back the spring to an angle and release it.

The vial strikes the platform, causing the flies to collide against the vial walls repeatedly.

The drosophila brain strikes against the head capsule, mimicking closed-head traumatic brain injury, or TBI, without causing external damage.

Allow the flies to recover, and transfer them to a vial with food.

The injury causes damage to neuron membranes, impairing neuronal function.

Signaling molecules from the damaged neurons recruit immune cells, leading to progressive degradation of brain tissue, eventually resulting in the flies' death.

Place between 1 and 60 carbon dioxide anesthetized flies in an empty vial, and stopper the vial using a tight fitting cotton ball. Then, draw a line on the vial, 1 inch from its base. Knock the flies down from the sides of the vial and push in the cotton ball until it matc