In Vivo Chamber Model
Categorized: Host-Pathogen Interactions
Day et.al., J. Infect. 2:39-51, 1980.
1. Preparation Of Chambers
A. Syringe Barrels:
- remove markings with 95% ethanol.
- use 1 ml syringes for mice & 3 ml syringes for rats.
- cut syringe barrels into 1 cm lengths.
- cut discs from 0.22 µm pore size millipore filters.
- use standard hole punch for mice & large 3-hole punch for rats.
- layer discs on paper in a glass petri dish.
- wrap well and autoclave.
2. Construction Of Chambers (in sterile hood)
- attach filter disc to one end of chamber with a little glue applied with a sterile toothpick.
- add 100 µl (for mice) or 500 µl (for rats) of bacterial sample (in buffer) to the chamber.
- add second filter to open end.
3. Implantation Of Chambers
- Anaesthetic: ketamine & xylazine mixed 1/10 in saline. Prepare 1/2 ml ketamine & 0.5 ml xylazine per mouse (amounts given per mouse listed in mouse room).
- Insert four chambers into the peritoneal cavity of each animal via a longitudinal incision in the abdomen.
- Close incision using surgical sutures.
4. Removal Of Chambers
- Kill animals with CO2.
- Chambers are removed by making an incision into the peritoneum.