IntelliProfiler 2.0: An integrated R pipeline for long-term home-cage behavioral profiling in group-housed mice using eeeHive 2D

BackgroundLong-term home-cage monitoring is essential to quantify spontaneous locomotor and social behaviors in group-housed mice, but analysis of high-density RFID tracking data remains a barrier to reproducibility. New methodsWe developed IntelliProfiler 2.0, a fully R-based pipeline tailored to the eeeHive 2D floor-mounted RFID array. The workflow performs data import from text logs, preprocessing, coordinate reconstruction, missing-value handling, feature extraction, statistical testing, and visualization in a single environment. Behavioral metrics include travel distance, close contact ratio (CCR), and a newly implemented inter-individual distance metric. ResultsIn four-day recordings of group-housed C57BL/6J mice (8 males and 8 females), IntelliProfiler 2.0 captured circadian phase-dependent locomotion and proximity patterns and reproduced sex-dependent differences consistent with prior analyses while incorporating updated hardware specifications. Radar-chart summaries enabled intuitive comparison of multidimensional behavioral profiles and inter-individual variability across light/dark phases. Comparison with existing methodsCompared with IntelliProfiler 1.0 and multi-tool workflows, IntelliProfiler 2.0 consolidates analysis into a single, script-based R pipeline, reducing operational complexity and improving reproducibility. The updated implementation supports recent manufacturer-driven changes, including antenna renumbering and multi-USB data export. ConclusionsIntelliProfiler 2.0 provides a reproducible, extensible framework for high-throughput behavioral phenotyping of group-housed mice and is scalable across hardware configurations, including simplified single-board recordings. HighlightsO_LIEnd-to-end R pipeline for eeeHive 2D floor-based RFID tracking analysis C_LIO_LIStandardized setup with comprehensive manuals and protocols C_LIO_LIInter-individual distance metric to quantify group spatial structure C_LIO_LICircadian- and sex-dependent behavioral profiling in group-housed mice C_LIO_LIRadar-charts summarize multidimensional behavioral profiles and variability C_LI