Generating a Mouse Model of Traumatic Brain Injury Using a Lateral Fluid Percussion Device

Take an anesthetized mouse and apply eye ointment to prevent its eyes from drying.

Shave the head, disinfect the scalp, and incise in the sagittal plane to expose the skull.

Remove the connective tissue layer and mark the target area for injury.

Drill to create a bone flap, then remove it to expose the dura mater, the brain's outer covering.

Place a cannula over the dura mater and secure it with dental cement.

Fill the cannula with saline, ensuring no air bubbles form, and connect it to the tubing of the lateral fluid percussion device, containing a pendulum and a fluid-filled reservoir.

Release the pendulum from a calibrated angle to strike the reservoir, generating a pressure pulse that travels through the fluid and reaches the brain.

The mechanical stress damages neurons, causing traumatic brain injury.

Remove the cannula, suture the incision, and allow the mouse to recover for further analysis.

For traumatic brain injury induction, confirm a lack of response to toe pinch in an anesthetized mouse, and apply ointment to the animal's eyes. After shaving, disinfect the exposed scalp skin with 70% ethanol before incising the scalp in a sagittal plane. Using forceps, retract the incision on both sides and separate the periosteum slightly.

Use a marker to draw a 3 millimeter diameter circle on the right parietal area of the skull, 2 millimeter away from the midline. Use an electric drill to carefully penetrate the skull without damaging the dura. Remove the bone flap to expose a 3 millimeter bone window, and place a plastic injury cannula over the craniotomy.

After cementing the cannula to the skull with dental acrylic, use a 5 milliliters syringe to fill the cannula with sterile 0.9% normal saline, taking care to avoid bubbles. Turn on the oscilloscope and amplifier. Confirm that the high pressure tube of the lateral fluid percussion entry device is free of air bubbles, and deliver about 10 test pulses until the device gives a steady signal.

Adjust the angle of the pendulum starting position to reach a pulse intensity of about two standard atmosphere, and connect the entry cannula to the lateral fluid percussion injury device. Pull the trigger to release the pendulum, inducing the brain injury. Then obtain and transmit a pulse to the dura through the entire closed fluid filled tubing system.

After brain injury induction, remove the cannula and suture the incision. Then, place the mouse on a heating pad with monitoring until it is ambulatory, before returning the animal to its home cage with ad libitum access to food and water.