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Protocol for non-invasive intratracheal instillation
The intratracheal instillation technique is used to deliver a variety of agents to the lungs. The intratracheal technique can involve the non-invasive instillation method or a direct injection into the trachea. In this protocol, we present the experimental procedure of the non-invasive instillation in the mouse as follows.
Anaesthetize the mouse by subcutaneous injection of ketamine (100 mg/kg) and xylazine (10 mg/kg).
Apply eye ointment such as Oculentum simplex, Teva Pharmachemie to the mouse eyes as to keep eyes moist.
The position of the intubation can be determined by dissecting a mouse before the experiment. In general, the reference distance for insertion is about 1.5 cm for a black adult mouse (25g).
Fix the anesthetized mouse on an inclined plane with an about 60° angle and suspend the mouse by its incisors with a piece of ribbon in the supine position on the platform, which will prevent the head from a violent shake.
Carefully open the mouse's mouth and gently pull the mouse tongue out with blunt-ended forceps. Insert the Exel Safelet IV indwelling needle (24G) slowly into the trachea, then gently remove the needle. Please note the distance is about 1.5 cm when inserting.
Inject 75 μl virus (or sterile PBS/normal saline as control) into the tip of the indwelling needle. Hold the mouse upright for a few seconds to allow virus to be inhaled into the lungs. It is necessary to control the volume of the virus so as to avoid the mouse being choked.
Post-injection care
Remove the mouse and place it prone on a heating pad for recovery.

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