Cranial Ring Assembly Installation in Mouse Model: A Surgical Procedure to Implant Visualization Window Assembly on Murine Skull for High-resolution Skull Bone Marrow Imaging

For cranial ring implantation on the mouse skull, first, prep an anesthetized mouse in the prone position. Next, make a midline vertical incision to expose the calvarium – the top part of the skull that protects the cranial cavity enclosing the brain.

Now, carefully remove the underlying periosteum — a dense layer of vascular connective tissue – and any tissue debris for optimal imaging. Clean the exposed skull surface with saline.

Next, apply cyanoacrylate glue to the metal skull ring. Mount it over the desired area of the dry exposed skull.

Hold the ring in place to allow it to attach firmly to the skull. Keep the skull surface hydrated with saline. Secure the ring to the skull area with dental cement to prevent any leakage during subsequent imaging.

Fill the ring with saline, which acts as the immersion medium for high-resolution microscopy.

Transfer the skull ring-bearing mouse to a support plate affixed to the block holder. The mouse’s head is raised to prevent ring detachment from the skull.

Fix the ring onto the block holder to immobilize the mouse’s head and prevent respiratory-induced motion artifacts during imaging. Position the support and the mouse assembly for intravital imaging through the skull bone, which facilitates the visualization of specialized cells and their native environment in real-time in the underlying bone marrow.