See JAX Pipeline on the IMPC website for order of tasks for all mice tested
Project protocol: Light-dark box test
Introduction
The light-dark box test measures the anxiety-related behavior of mice in a two-chambered light-dark arena.
Study design
Paradigm: Baseline study
Panel: Collaborative Cross
Number of strains: 18
Sex: Both
Age: 8-10 wks
Sample size: Seven mice per sex from at least two litters
Number of cohorts: 1
Workflow
Step |
Description |
Equipment |
Data collected |
1 |
Mice are acclimated to test room for at least 3 days |
- |
- |
2 |
Each mouse is placed in the light side of the arena |
Light-dark box |
- |
3 |
Behavior is recorded for 20 min |
Light-dark box |
Latency to enter dark zone, time mobile in each zone, time spent in each zone, percentage of time in each zone, number of transitions between zones, number of fecal boli |
Equipment, software and supplies
Reagents and solutions
Procedure
- Mice are allowed to acclimate to the testing room for a minimum of 3 days.
- Light levels within the room are set so that the light intensity in the light arena is between 100-200 lux and 4-7 lux in the dark arena. Light metering is performed at the start of each test day.
- Photobeam sensor check is performed using the built-in diagnostic function at the beginning and end of every trial.
- Each mouse is placed in the light side of an arena.
- Behavior is recorded for 20 min.
- At the end of the 20 min test, mice are removed from the arenas and returned to their home cages.
- Arenas are cleaned with 1% Virkon S solution to disinfect and remove scent cues from previously tested mice.
- Analysis of the recording is done to measure the activity of each mouse in each of the two zones and the number of transitions between zones.
Data collection: The Accuscan Versamax system in configured to report parameters for each mouse. Versamax files are converted into text data files using Versadat. It outputs a .DPP file containing light-dark information. Raw data is stored until outlier detection and other QA/QC procedures are complete. Local data processing is implemented to calculate derived measures from this data and a table of the derived measures and raw time per bin per zone is reported per mouse in the LIMS system.
Data QC: Verify that total time of each run is 20 min.
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 zone are reported per mouse in JaxLIMS.
Measurements
LIMS field name |
MPD variable name |
Description |
Units |
Series |
Primary measurement |
Series order |
MPD computed |
Calculation |
Method |
Data type |
Side changes |
transitions_LD |
number of transitions between light and dark zones, 20 min test |
n |
|
|
|
|
|
light-dark box test |
Integer |
Left side time spent |
light_dur_LD |
time spent in light zone, 20 min test |
s |
|
|
|
|
|
light-dark box test |
Float |
Time mobile left side |
time_mobile_LD |
time mobile in light zone, 20 min test |
s |
|
|
|
|
|
light-dark box test |
Float |
Right side time spent |
dark_dur_LD |
time spent in dark zone, 20 min test |
s |
|
|
|
|
|
light-dark box test |
Float |
Time spent mobile right side |
time_mobile_LD |
time mobile in dark zone, 20 min test |
s |
|
|
|
|
|
light-dark box test |
Float |
Fecal boli |
defecation_LD |
fecal boli count, 20 min test |
n |
|
|
|
|
|
light-dark box test |
Integer |
Reaction time |
reaction_time_LD |
latency to enter dark zone, 20 min test |
s |
|
|
|
|
|
light-dark box test |
Float |
Percent time in dark |
pct_dark_LD |
percentage of total time in dark zone, 20 min test |
% |
|
|
|
|
|
light-dark box test |
Float |
Percent time in light |
pct_light_LD |
percentage of total time in light zone, 20 min test |
% |
|
|
|
|
|
light-dark box test |
Float |
Experimental and technical parameters
Light level in light zone |
100-200 lux |
Light level in dark zone |
4-7 lux |
Equipment name |
Light-Dark Insert |
Equipment manufacturer |
Accuscan |
Equipment model |
Accuscan Versamax |
Start time |
0700 - 1700 EST |
Disinfectant |
1% Virkon Solution |
|