Performing Intratracheal Bacterial Delivery in a Mouse Model

Samenvatting

Source: Venuprasad, K., et al. Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation. J. Vis. Exp. (2016)

This video demonstrates the intratracheal delivery of a lung-pathogenic bacterial inoculum into an anesthetized mouse to model lower respiratory tract infection.

Protocol

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

1. 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.
2. Pinch the interdigital space of the mouse to verify that the mouse is adequately anesthetized.
3. Place the mouse on its back. Shave the ventral area of the neck and disinfect with chlorhexidine and 70% ethyl alcohol.
4. 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.
   1. 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.
5. Close the incision with surgical glue. Keep the mouse laterally recumbent and warm for about 10 min.

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
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