Spinal Cord Window Implantation in a Mouse for In Vivo Imaging

Begin with an anesthetized mouse with a surgically exposed spinal cord.

Place a borosilicate cover glass over the spinal cord to create an optical window for imaging.

Apply warm agarose to the edges of the cover glass, allowing capillary action to draw it underneath to maintain tissue hydration.

Introduce tissue adhesive to the intact bony structures surrounding the site to stabilize the spinal cord window.

Apply dental cement mixed with an accelerant over the adhesive layer, then position a 3D-printed backplate over the cover glass before the cement cures.

Apply additional thin layers of cement to strengthen backplate adhesion.

After the cement has dried, secure the backplate to a forked backplate holder to minimize motion artifacts from respiration and heartbeat.

Add saline to check for leakage and seal with additional cement if necessary.

Finally, transfer the mouse and platform to a two-photon microscope for imaging.

For cover glass implantation, gently apply a 3 millimeter borosilicate cover glass to the exposed cord. Use a small spatula to apply 39 degrees Celsius, warmed 2% agarose to the edge of the glass, and allow capillary action to draw the agarose under the cover glass surface