Intrabronchial Delivery: A Technique to Administer an Experimental Agent Selectively Into a Mouse Lung
Transcrição
– An intrabronchial delivery administers an experimental agent to a particular lung. First, fix an anesthetized mouse to a plastic board in a supine position, and cannulate the trachea with an extended catheter. To cannulate the distal right lung, rotate the board to 30 degrees and guide the catheter parallel to the mouse's midline until you feel resistance.
Administer the experimental agent into the catheter. Then with a dropper, dispense air to clear the agent from the catheter. Withdraw the catheter and maintain the mouse in position for 30 seconds. Place the mouse in a cage for recovery.
To deliver experimental agents to the distal left lung following intratracheal cannulation, rotate the plastic board to minus 74 degrees. Gently advance the catheter into the left mainstem bronchus. Rotate the board to minus 30 degrees to allow gravitational assistance for experimental agent administration. Deliver more volume of the experimental agent proportionate to the left lung lobe having a larger area. With a dropper, dispense air and withdraw the catheter. Maintain the mouse in position for 30 seconds, then place it in a cage to recover.
The following protocol shows Evans blue dye delivery via intrabronchial administration for selective deposition in a mouse lung.
– For selective lobar cannulation of the distal right lung, after intratracheal intubation, rotate the plastic board 30 degrees, and guide the hub of the catheter in parallel to the mouse midline to the appropriate weight based depth as indicated in the table. Using a gel loading tip, deliver 20 microliters of 0.3% Evans blue dye into the catheter followed by 1 to 2 100 microliter aliquots of air by glass dropper. Then withdraw the catheter and maintain the mouse in position for 30 seconds before placing the animal on a warming blanket with monitoring until full recovery.
For selective lobar cannulation of the distal left lung, after intratracheal intubation, rotate the plastic board minus 74 degrees and gently advance the catheter into the left main stem bronchus while placing modest pressure at 90 degrees downward and toward the bookend. After cannulating the left lower lung segments, rotate the plastic board minus 30 degrees and deliver 40 microliters of 0.3% Evans blue dye with a gel loading tip. Then deliver 1 to 2 100 to 300 microliter aliquots of air via glass dropper and withdraw the catheter to allow the mouse to recover as demonstrated.
