Convection-Enhanced Delivery of Antibodies into a Mouse Brain
Transcripción
For antibody injection into the murine striatum, confirm a lack of response to skin pinch in an anesthetized mouse, and shave the head with a hair trimmer. Disinfect the skin with cotton swabs soaked in iodine solution.
Use a scalpel to make a 10-millimeter skin incision along the cranial midline finishing at the eye level. Fix the mouse in the stereotactic frame using the nose clamp and ear bars, taking care that the skull surface is horizontal and tightly secured.
Place the syringe in the stereotactic robot, and synchronize the drill bit with the tip of the catheter on a reference point. Use forceps to retract the skin, and localize bregma on the skull surface. Reference bregma in the software using the tip of the drill bit, and move the drill to a 1-millimeter frontal and 2-millimeter lateral from bregma position.
Drill a burr hole, taking care not to damage the dura mater, and move the syringe over the burr hole. Dispense 0.5 to 1 microliter from the syringe to ensure that no air bubbles are left in the catheter. Start the convection-enhanced delivery as demonstrated, observing the skull surface for any traces of fluid backflow from the injection spot.
At the end of the delivery and catheter removal, start the injection pump at 0.2 microliters per minute to check for evidence of catheter clogging during the injection. If no clogging occurred, a droplet of injection mix should immediately be observed coming from the catheter tip.
