This article describes a method for directly delivering lung-pathogenic bacteria into the lower respiratory tract of anesthetized mice. The technique allows for bypassing upper airway barriers, facilitating the study of respiratory infections.
Begin with an anesthetized mouse in the supine position, with its neck shaved.
Disinfect the shaved area.
Make an incision in the neck, then dissect the muscle to expose the trachea, the airway leading to the lower respiratory tract, including the lungs.
Take a syringe pre-loaded with lung-pathogenic bacteria, positioned between two small air volumes.
Insert the needle into the trachea and inject the first air volume to clear the trachea.
Next, deliver the bacterial inoculum, followed by the second air volume to enhance the bacterial dispersion deeper into the lower respiratory tract.
Briefly hold the mouse vertically to promote an even distribution of bacteria throughout the lower respiratory tract.
Seal the incision with tissue adhesive and allow the mouse to recover.
This method enables direct delivery of pathogens into the lower respiratory tract, bypassing the upper airway barriers.
Begin with anesthetizing the mouse, and verifying it is adequately anesthetized using a toe pinch.
Then, position the mouse supine and shave the ventral area of the neck. Disinfect the exposed skin. Now, lift the skin of the upper ventral part of the neck with rat tooth forceps, and using a scalpel, make a one half to one centimeter incision above the thymus.
Then, carefully separate the muscle to expose the trachea. Into the trachea, slowly inject 50 microliters of air, followed by 50 microliters of prepared NTHi, followed by another 50 microliters of air.
After the injection, keep the animal vertical for around a minute. Then, close the incision with tissue adhesive and keep the mouse laterally recumbent and warm for about 10 minutes.
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