The Open Field test is used to assess anxiety and exploratory behaviors. It is based on the natural tendency of an animal to explore and to protect itself using avoidance which translates to a normal animal spending more time in the periphery of the Open Field arena than in the center (the most anxiogenic area).
- Minimum number of animals : 7M + 7F
- Age at test: Week 9
- Sex: We would expect the results of this test to show sexual dimorphism
1. Animals are transported to the testing room (or ideally an antechamber) and left undisturbed for 30 minutes before the test.
2. Ensure that lighting conditions are as desired (150-200 lux) and that all equipment is working correctly:
- If using a beam break system, ensure that beams are not interrupted before putting the mouse and if measuring vertical activity (rears), check that height of the infrared frame for rears is correctly set.
- If using a videotracking system, set detection parameters appropriate for the coat color of the mice.
- For analysis, each openfield arena is divided into a peripheral zone measuring 8 cm from the edge of the arena walls, and a central zone around 40% of the total surface of the arena.
3. Testing is conducted during the light phase of the cycle with 1 hour gap from the light/dark change. Testing should be conducted during the same period of day.
4. Wipe the apparatus clean and allow time for it to dry
5. Each mouse is placed in the middle of a peripheral zone of the arena facing the wall and allowed to explore freely the apparatus, with the experimenter out of the animal's sight.
If more than one mouse can be tested in parallel, in adjacent open field arenas and mice are video-tracked, it is important to ensure that the tracking of each mouse starts as soon as the mouse is released to make data comparable.
Males and females must be run in separate tests; ideally males are tested first, then followed by females
6. At the end of the 20 min run, animals are labeled (if necessary) and put back into their home cage.
7. After each run, any feces are removed and the arena is thoroughly wiped.
8. Analysis of the recording is done to measure the activity of each mouse in each of the zones per 5 minute bins.
- Please ensure that the mice are not handled prior to the test, except handling for cage change.
- Ear clipping should be done after the test if necessary .
- Verify total time of run.
- Coherence between latency, number of entries and time in the center; e.g. it is not possible to have zero entries and have spent some time spent in the center.