Implantation of Cranial Imaging Window in Mouse Model: A Surgical Procedure to Implant Glass-based Coverslip for Stable Optical Access to Regions of Murine Brain

After confirming a lack of response to toe pinch in an anesthetized adult mouse, mount the mouse on a stereotactic frame and secure the head with a nose clamp and two ear bars.

Use a heating lamp to preserve the body temperature and apply ointment to the animal's eyes. Using sharp scissors, shave the fur on the skull from the eyes to the base of the skull, and use 70% ethanol to sterilize the exposed skin.

Cut the skin in a circular manner, and use a cotton swab to scrape away the exposed periosteum. Treat the surgical area with a drop of 1% lidocaine and a 1:100,000 concentration of epinephrine for 5 minutes before removing the excess solution with a cotton swab.

Use cyanoacrylate glue to adhere the edges of the skin to the skull, and place the stereotactic frame under a dissection stereomicroscope with a 4X magnification.

Next, carefully drill a superficial 5-millimeter diameter circular groove over the right parietal bone and apply a drop of cortex buffer to the groove.

Use thin forceps to lift the bone flap to visualize the brain surface, and use curved tapered, very fine point forceps to remove the dura mater. If bleeding occurs, use an absorbable gelatin sponge to achieve hemostasis.

To prepare the cranial imaging window, replace the cortex buffer with a drop of silicone oil to the craniotomy site to avoid air bubbles under the window.

Seal the exposed brain with a 6-millimeter coverslip and apply cyanoacrylate glue between the coverslip and the skull. Then, use fine tweezers to gently press the coverslip against the skull.