Purpose
This SOP defines standardized execution, scoring, and documentation for the Open Field in mouses.
Plan, acclimate, score, document, and run rodent behavior batteries with five free browser tools.
Pick tests, then lint for confounded orders, one-shot repeats, stress overload, and recovery-window mismatch.
1. Open Field
Recovery after: 1 day | one-shot: no
2. Rotarod
Recovery after: 1 day | one-shot: no
3. Elevated Plus Maze
Recovery after: 1 day | one-shot: yes
4. Forced Swim Test
Recovery after: 7 days | one-shot: yes
No major sequencing warnings detected.
| Day | Test | Paradigm | Stress | Links |
|---|---|---|---|---|
| 1 | Open Field | anxiety | 1 | Calculator/Apparatus |
| 2 | Rotarod | motor | 2 | Calculator/Apparatus |
| 3 | Elevated Plus Maze | anxiety | 2 | Calculator/Apparatus |
| 4 | Forced Swim Test | depression | 5 | Calculator/Apparatus |
| Day | Date | Activity | Cohort | Details |
|---|---|---|---|---|
| -7 | 2026-04-24 | Room habituation begins | All cohorts | Move animals into the test suite or holding room at least 7 days before testing. |
| -6 | 2026-04-25 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| -5 | 2026-04-26 | Handling sessions | Cohort A | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -5 | 2026-04-26 | Handling sessions | Cohort B | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -5 | 2026-04-26 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| -4 | 2026-04-27 | Handling sessions | Cohort A | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -4 | 2026-04-27 | Handling sessions | Cohort B | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -4 | 2026-04-27 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| -3 | 2026-04-28 | Handling sessions | Cohort A | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -3 | 2026-04-28 | Handling sessions | Cohort B | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -3 | 2026-04-28 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| -2 | 2026-04-29 | Handling sessions | Cohort A | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -2 | 2026-04-29 | Handling sessions | Cohort B | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -2 | 2026-04-29 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| -1 | 2026-04-30 | Handling sessions | Cohort A | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -1 | 2026-04-30 | Handling sessions | Cohort B | Three 1 minute handling sessions per animal, separated by at least 30 minutes. |
| -1 | 2026-04-30 | Room habituation | All cohorts | Keep husbandry, transport, noise, and room entry windows consistent. |
| 0 | 2026-05-01 | Begin behavior battery | Cohort A | Run Cohort A at the same circadian phase and document transport order. |
| 1 | 2026-05-02 | Begin behavior battery | Cohort B | Run Cohort B at the same circadian phase and document transport order. |
Walking
Locomotion with four paws, head forward.
Supported rear
Rearing with forepaws on a wall or object.
Body grooming
Licking or scratching trunk, flank, tail, or hindlimbs.
Sniffing conspecific
Nose-to-nose, nose-to-body, or nose-to-anogenital contact.
Freezing
Complete immobility except respiration for at least 1 second.
Cohen's kappa
0.545
Moderate agreement
Interpretation follows Landis and Koch categories.
This SOP defines standardized execution, scoring, and documentation for the Open Field in mouses.
Use Open Field Arena. Confirm apparatus dimensions, camera coverage, software project settings, and calibration records before the first animal is tested.
Test mouses after the approved acclimation and handling period. Maintain consistent transport order, holding conditions, and circadian timing across cohorts.
Record lux at the arena floor before each testing block.
Clean with 70% ethanol between animals and allow surfaces to dry.
Run Open Field according to the approved protocol. Record animal ID, group, cohort, start time, apparatus ID, software version, operator, and any deviations.
- Tracking failure that prevents primary endpoint scoring - Animal escape from apparatus - Protocol deviation documented before unblinding
Score primary endpoints from blinded video files. Record start time, endpoint values, and tracking quality.
Compare predefined groups using the approved model. Report n, exclusions, effect size, and confidence interval.
Crawley 2007; McIlwain et al. 2001; Bailoo et al. 2014; local IACUC protocol and lab SOP index.
Every test in the catalog links to its dedicated calculator at /tools/<test-name> and apparatus at /lab/<apparatus>. For automated tracking, see ConductVision.
Don't use for
A behavior battery is not just a list of assays. Each test can change arousal, fatigue, learning, pain sensitivity, social motivation, or anxiety-like behavior. Low-stress exploration and baseline motor checks usually belong early. Aversive or one-trial tests need careful placement because they can create carryover effects that are larger than the biology being measured.
Use the sequencer as a planning pass before calendar lock. The warnings do not replace protocol review, but they make common order effects visible while the schedule is still easy to change.
Acclimation is an experimental variable. Room transfer, reversed light cycles, handling, cage change timing, and transport order can all create cohort effects. A useful schedule names the day, cohort, activity, and handling dose so technicians can run the same preparation every time.
The tracker starts from the battery date and works backward. That keeps the first test from becoming the only fixed point while animal preparation drifts.
Behavior scoring becomes reliable when raters share operational definitions. Ethograms convert vague labels into observable states, and IRR checks reveal where raters disagree before the main dataset is scored.
For categorical states, Cohen kappa is a compact check on agreement beyond chance. For continuous measurements, ICC(2,1) estimates absolute agreement between raters on the same subjects.
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