Three-Chamber Social Novelty Preference
Overview
The three-chamber social novelty preference test constitutes Phase 2 of the Crawley sociability paradigm, designed to assess social recognition memory and preference for novelty in a familiar versus unfamiliar social context. Following the initial sociability phase where the subject interacts with a single stranger mouse (S1), a second novel stranger (S2) is introduced into the previously empty wire cage on the opposite side of the apparatus. The test subject, which remains in the center chamber during the stimulus changeover, is then given free access to both side chambers for a 10-minute exploration period. This phase specifically isolates social discrimination by requiring the subject to distinguish between a recently encountered conspecific and an unfamiliar individual.
The primary dependent variable is the social novelty preference index, calculated as (time near S2 minus time near S1) divided by (time near S2 plus time near S1), yielding a score from negative one to positive one where positive values indicate preference for the novel stranger. Typically developing C57BL/6 mice show preference indices between 0.15 and 0.40, while models of autism spectrum disorder such as BTBR and Shank3-knockout mice display indices near zero or negative values, reflecting impaired social recognition. Additional measures include direct sniffing bouts directed at each wire cage, the number of transitions between chambers as an index of general exploratory motivation, and the discrimination ratio computed from sniffing durations rather than proximity time.
ConductMaze tracks the subject animal in real time across the three-chamber apparatus using overhead video with center-point and nose-point detection, automatically computing proximity time within predefined interaction zones surrounding each wire cage. The system logs chamber entries, time spent in each chamber, and cumulative sniffing duration using nose-point distance thresholds calibrated to the wire cage dimensions. Automated preference index calculations are generated immediately after each trial, and batch analysis across cohorts produces group-level social novelty scores with built-in statistical comparisons between S1 and S2 interaction metrics.
Trial Flow
Apparatus Setup
Confirm three-chamber apparatus with wire cages in each side chamber; verify removable partitions are functional
Stimulus Placement
Familiar stranger S1 remains in original wire cage; introduce novel stranger S2 into opposite wire cage
Subject Release
With subject in center chamber, simultaneously remove both partition doors to allow free access
Free Exploration
Subject freely explores all three chambers for the test duration while video tracking records position
Proximity Detection
Nose-point within 2 cm of wire cage is scored as active investigation; center-point within interaction zone as proximity time
Chamber Classification
Each position frame is classified by chamber (S1 side, center, S2 side) using partition boundaries
Preference Computation
Calculate social novelty preference index, discrimination ratio, chamber times, and transition counts
Trial End
Remove subject and stimulus mice; clean apparatus and wire cages with 70% ethanol between cohorts
Parameters
| Parameter | Type | Default | Description |
|---|---|---|---|
| Test Duration | duration | 600 | Total exploration time for Phase 2 in seconds (standard 10 min) |
| Habituation Duration | duration | 300 | Phase 1 sociability phase duration in seconds (completed prior) |
| Interaction Zone Radius | distance | 5.0 | Radius around wire cage defining the interaction zone in centimeters |
| Sniffing Threshold | distance | 2.0 | Maximum nose-point to wire cage distance scored as active sniffing in centimeters |
| Chamber Width | distance | 20.0 | Width of each chamber in centimeters (standard 20 cm for mice) |
| Chamber Length | distance | 40.0 | Total apparatus length across all three chambers in centimeters |
| Partition Door Width | distance | 5.0 | Width of the opening between chambers in centimeters |
| Illumination | integer | 25 | Ambient illumination level in lux (low light reduces anxiety confounds) |
| S1 Habituation Time | duration | 300 | Time S1 spent in apparatus during Phase 1 in seconds |
| Stimulus Strain | enum | age-matched | Strain and sex match criteria for stimulus mice (age-matched, same-sex) |
Metrics
| Metric | Unit | Description |
|---|---|---|
| Social Novelty Preference Index | index | (S2 time - S1 time) / (S2 time + S1 time); range -1 to +1 |
| Time Near S2 | seconds | Total time spent in the interaction zone around the novel stranger wire cage |
| Time Near S1 | seconds | Total time spent in the interaction zone around the familiar stranger wire cage |
| S2 Sniffing Duration | seconds | Cumulative nose-point investigation time directed at the novel stranger cage |
| S1 Sniffing Duration | seconds | Cumulative nose-point investigation time directed at the familiar stranger cage |
| Chamber Transitions | count | Total number of crossings between chambers during the test |
| Center Chamber Time | seconds | Time spent in the center (neutral) chamber |
| Latency to S2 | seconds | Time from door opening to first entry into the S2 interaction zone |
Sample Data
| Subject | Genotype | Preference Index | S2 Time (s) | S1 Time (s) | S2 Sniffing (s) | Transitions |
|---|
Representative data for illustration purposes. Actual values will vary by species, strain, and experimental conditions.
Applications
- 1Autism spectrum disorder modeling — detecting social recognition deficits in Shank3, CNTNAP2, and BTBR mouse models
- 2Oxytocin pharmacology — evaluating prosocial drug candidates that rescue social novelty preference in deficit models
- 3Social memory circuits — dissecting hippocampal CA2 and medial amygdala contributions to familiar-novel discrimination
- 4Neurodevelopmental screening — high-throughput phenotyping of social cognition across transgenic lines
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