Cranial window implantation is a surgical technique that allows for long-term in vivo imaging of the brain. This procedure involves creating a window in the skull and sealing it with a coverslip to facilitate access to the brain.
Cranial window implantation is a surgical procedure wherein an exposed region of the brain is sealed with a coverslip. The implanted imaging window facilitates access to the brain for long-term high resolution in vivo imaging.
To begin, stabilize an anesthetized mouse in a stereotactic unit.
Maintain its body temperature using a heating pad.
Next, shave the mouse’s scalp and sterilize the exposed skin.
Make a circular incision and scrape off the underlying periosteum - a fibrous connective tissue layer - from the skull surface.
Subsequently, apply a suitable vasoconstrictor and an anesthetic solution over the surgical area to minimize bleeding in the following steps.
Secure the skin edges to the skull with cyanoacrylate glue.
Then, under a dissection microscope, visualize the skull.
Drill a shallow groove over the region of interest until the bone flap becomes loose. Treat the groove with cortex buffer to prevent tissue dehydration.
Gently lift the bone flap, creating a bone window to access the brain underneath.
Now, remove the outermost membrane layer surrounding the brain - the dura mater. Clear the buffer and apply silicone oil to prevent air bubbles under the window.
Mount a coverslip over the exposed area using glue, ensuring minimal distance between the brain and the coverslip.
The mouse with the implanted cranial window is ready for brain imaging.
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.
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