Procedure:
Acclimation (5 min)
a. Make sure the confinement cylinders are on the left and right sides of the test arena according to
the zone map.
b. Insert the sliding doors in each dividing wall, thereby blocking access of the left and right
chambers.
c. Place the subject mouse in the center arena and allow acclimation for 5 minutes.
Trial 1 Social Affiliation (10 min)
a. Following acclimation, retrieve 1 control mouse (stranger 1) from holding cage and place in the
left cylinder. Right cylinder remains empty.
b. Remove the sliding doors and immediately click once on “Start” to begin video tracking.
c. Click the red record button to start recording the video.
d. Leave the test room and make sure the door is closed. Monitor the recording session patiently
and quietly.
e. Once the trial is over, manually stop the video by hitting the same record button andclick the
save button at the top of the program screen to save data.
f. Return subject mouse to the center of the arena and replace the sliding doors.
Trial 2 Social Novelty (10 min)
a. Following trial 1, retrieve the second control mouse (stranger 2) from holding cage and place in
the right cylinder. The first mouse (stranger 1) remains in the left cylinder.
b. Retrieve the subject mouse and place in the middle chamber of the test arena.
c. Remove the sliding doors and immediately click once on “Start” to begin video tracking.
d. Click the red record button to start recording the video. When the run is over, manually stop the
video by hitting the same record button.
e. Leave the test room and make sure the door is closed. Monitor the recording session patiently and quietly.
Clean-up
a. Return all mice to their respective home cage.
b. Clean the test arena and cylinders with 70% ethanol then with wipe down with nolvasan. Repeat the process for subsequent mice.