JaxCC1 project protocol for procedure: open field

Multisystem phenotyping of 18 Collaborative Cross strains   (2018)

The Jackson Laboratory


Open field test


The open field test monitors the activity of mice in an open arena over a defined period of time in order to assess locomotor activity, anxiety, exploratory behavior, and habituation phenotypes.

Study design

  • Paradigm: Baseline study
  • Panel: Collaborative Cross
  • Number of strains: 18
  • Sex: Both
  • Age: 7-9 wks
  • Sample size: Seven mice per sex from at least two litters
  • Number of cohorts: 1


    Step Description Equipment Data collected
    1 Mice are acclimated to test room for at least 3 days - -


    Each mouse is placed in the center of an arena Open field arena -
    3 Behavior is recorded for 20 min Open field arena Locomotor activity, anxiety- and exploratory-related behaviors

    Equipment, software and supplies

    Reagents and solutions

    • 1% Virkon S solution



    1. Mice are allowed to acclimate to the testing room for a minimum of 3 days.
    2. Light levels within the room are set so that the light intensity in each arena is between 100-200 lux. Light metering is performed at the start of each test day.
    3. Photobeam sensor check is performed at the start of each test day or in the event of drift in control parameters using the built-in diagnostic function.
    4. The method of recording is:
      • Peripheral zone: 7.62 cm from the edge of the arena walls
      • Center zone: ~40% of the total surface area of the arena
      • Corner zones: 58.06 cm2 each
    5. Each mouse is placed in the center of each arena, nose facing to experimenter's left and released.
    6. After all mice have been placed within an arena, behavior is recorded for 20 min.
    7. At the end of the 20 min test, mice are removed from the arenas and returned to their homecages.
    8. Arenas are cleaned with 1% Virkon solution to disinfect and remove scent cues from previously tested mice.
    9. Analysis of the recording is done to measure the activity of each mouse in each of the three zones in 5-min bins.

    Data collection: Data are reported as an activity summary in both seconds moving and cm per bin and zone. Versamax files are converted into text data files using the programs Versadat and Versamap. The resulting output is a .ACT file (containing general movement information) and Versamap (containing zone information).

    Formulas and derivations: Local data processing is implemented to calculate derived measures from these data, and a table of the derived measures and raw time per bin per zone are reported per mouse in JaxLIMS.


    Experimental and technical parameters

    Light level 100-200 lux
    Central zone surface area Total area is 645.16 cm2 and 39.06% of whole arena
    Peripheral zone surface area 7.62 cm from wall (total area is 1006.45 cm2 and 60.94% of whole arena)
    Corner zone surface area Total area is 232.26 cm2 and 14.06% of whole arena
    Arena size 42 x 42
    Equipment name Open Field
    Equipment manufacturer Accuscan
    Equipment model Accuscan Versamax (VMX 1.4b)
    Color of arena Grey
    Configuration file/software version Versamax 4.2
    Type of analysis Wide beam break
    Start time 0700 - 1700 EST
    Disinfectant 1% Virkon Solution