In Vivo Chamber Model

Reference:

Day et.al., J. Infect. 2:39-51, 1980.

1. Preparation Of Chambers

A. Syringe Barrels:

1. remove markings with 95% ethanol.
2. use 1 ml syringes for mice & 3 ml syringes for rats.
3. cut syringe barrels into 1 cm lengths.

B. Filters:

1. cut discs from 0.22 µm pore size millipore filters.
2. use standard hole punch for mice & large 3-hole punch for rats.
3. layer discs on paper in a glass petri dish.
4. wrap well and autoclave.

2. Construction Of Chambers (in sterile hood)

1. attach filter disc to one end of chamber with a little glue applied with a sterile toothpick.
2. add 100 µl (for mice) or 500 µl (for rats) of bacterial sample (in buffer) to the chamber.
3. add second filter to open end.

3. Implantation Of Chambers

1. 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).
2. Insert four chambers into the peritoneal cavity of each animal via a longitudinal incision in the abdomen.
3. Close incision using surgical sutures.

4. Removal Of Chambers

1. Kill animals with CO_2.
2. Chambers are removed by making an incision into the peritoneum.