All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.1. Intra-tracheal (i.t.) Instillation<ol><li>Inject the mouse intraperitoneally (i.p.) with 0.1 mL/10 g body weight of mouse with a 150 mg/ml ketamine + 20 mg/ml xylazine solution. The procedure should be done in the hood with a light source to keep the animal warm.</li><li>Pinch the interdigital space of the mouse to verify that the mouse is adequately anesthetized.</li><li>Place the mouse on its back. Shave the ventral area of the neck and disinfect with chlorhexidine and 70% ethyl alcohol.</li><li>Lift the skin of the upper ventral part of the neck using rat tooth forceps. Make a small incision (0.5-1.0 cm) above the thymus using a scalpel and carefully dissect the muscle to expose the trachea.<ol style='list-style-type:decimal;'><li>Using a 1 ml syringe with a 27 G needle, slowly inject 0.05 ml air, followed by 0.05 ml non-typeable Haemophilus influenzae (NTHi) or saline as a control, followed by another 0.05 ml air. Keep the animal vertical for around 1 min.</li></ol></li><li>Close the incision with surgical glue. Keep the mouse laterally recumbent and warm for about 10 min.</li></ol>

<table><tbody><tr><td>Deft quick solution</td><td>Sigma</td><td>GS500-500ML</td></tr><tr><td>Syringe needle 20/26 G</td><td>BD</td><td>REF305115/175</td></tr><tr><td>1 mL syringe</td><td>BD</td><td>REF 309602</td></tr><tr><td>Dissecting scissors, straight, 10 cm long</td><td>Kentscientific</td><td>INS600393</td></tr><tr><td>Iris forceps, serrated, 10 cm long</td><td>Kentscientific</td><td>INS650915</td></tr><tr><td>Tweezer 5 stainless steel, 11 cm long</td><td>Kentscientific</td><td>INS600095</td></tr></tbody></table>

performing intratracheal bacterial delivery in a mouse model

Performing Intratracheal Bacterial Delivery in a Mouse Model

Source: <a href='https://app.jove.com/v/53964/intra-tracheal-administration-haemophilus-influenzae-mouse-models-to'>Venuprasad, K., et al. Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation. J. Vis. Exp. (2016)</a>This video demonstrates the intratracheal delivery of a lung-pathogenic bacterial inoculum into an anesthetized mouse to model lower respiratory tract infection.

Source: Venuprasad, K., et al. Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation. J. Vis. Exp. ...

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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746 Views. 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 bacter...

Performing Intratracheal Bacterial Delivery in a Mouse Model

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Tags

Name	Company	Catalog Number
Deft quick solution	Sigma	GS500-500ML
Syringe needle 20/26 G	BD	REF305115/175
1 mL syringe	BD	REF 309602
Dissecting scissors, straight, 10 cm long	Kentscientific	INS600393
Iris forceps, serrated, 10 cm long	Kentscientific	INS650915
Tweezer 5 stainless steel, 11 cm long	Kentscientific	INS600095