Visualizing Cerebral Blood Flow During Ischemia in a Mouse Using a Fluorescent Tracer

Secure an anesthetized mouse in a supine position.

Make a neck incision to expose and loosely ligate the bilateral common carotid arteries.

Loosely close the incision and position the mouse in a prone position.

Shave the scalp, disinfect, and make an incision to expose the skull.

Remove the soft tissue and glue a metal frame to the skull over the region of interest.

Secure the mouse on a steel plate.

Drill to thin the skull until surface cerebral blood vessels become visible.

Inject fluorophore-tagged dextran into the tail vein.

Dextran circulates through the bloodstream and reaches the brain, labeling the cerebral blood vessels.

Using a fluorescence microscope, visualize cerebral blood flow through the thinned skull.

Position the mouse supine and tighten the carotid sutures to restrict cerebral blood flow, simulating ischemia.

Microscopically re-visualize through the thinned skull to identify ischemic areas with reduced blood flow and detect plasma leakage.