Figure 4.

Surgeries for implanting terminal windows in the mouse brain

Left panel: Preparation of surgical site. Begin by injecting anesthetic intraperitoneally. Diagram and picture show site of incision. Three options exist for surgical preparation: a) Option 1: No skull thinning. Coronal view shows untouched cranium and meninges. b) Option 2: Skull thinning using surgical drill. Coronal view shows thinned cranium with untouched meninges. c) Option 3: Surgical creation of cranial window. This involves the removal of the skull cap, and imaging directly on the brain. Coronal view shows full view into the brain. Middle panel: Place window for imaging. Several options exist for imaging. Option 1: no coverslip is placed, and imaging is done directly on the hydrated exposed tissue. Option 2: A round coverslip is placed using surgical glue. Option 3: A round coverslip attached to a vacuum ring is placed on top of the incision. Right panel: Place mouse on stage and image. The head of the mouse must be secured/immobilized using a stereotaxic instrument. This includes securing at 3 main points: the teeth, and both ears. A support for the chin can be used to help immobilization. Depending on the experimental conditions, dyes can be added intravenously either before or after securing the mouse on the stage. An upright microscope is best suited to image the brain. The objective reaches the brain from the top. Scale bar: 20 μm. Created with BioRender.com.