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eeeHive: a new HF RFID-based automated behavioral monitoring system for group-housed animals with high spatiotemporal resolution

Authors: Seico Benner (NIES Tsukuba; Hamamatsu University School of Medicine), Suzuka Shiono (Phenovance LLC), Toshinori Kagawa (CRIEPI), Kiyohiko Hattori (Tokyo Denki University), Hidenori Yamasue (Hamamatsu), Hans-Peter Lipp (University of Zurich), Toshihiro Endo (Phenovance LLC). Preprint: bioRxiv 2026.04.30.720993 (doi 10.64898/2026.04.30.720993). Not peer reviewed.

Abstract

Long-term, automated tracking of group-housed social animals using RFID (radio frequency identification) is a promising approach in ethological neuroscience. However, low-frequency (LF) RFID, while long-established in the field, is constrained by its inherent low data rates, which lead to two critical limitations: ( 1) compromised spatiotemporal resolution, and ( 2) the inability to identify multiple tags (animals) simultaneously. To address these limitations, we developed eeeHive, a high-frequency (HF) RFID-based animal tracking system with a fully custom hardware architecture that enables high-speed, multiplexed antenna polling and concurrent multi-tag reading. The polling time per antenna in eeeHive was 5.9 ms, with an additional 8.2 ms read time per tag. We applied the system to track 24 mice for one week, and six common marmosets for seven weeks. The system successfully tracked individuals even within dense clusters, revealing complex behavioral traits characterized by spatial utilization, temporal dynamics, behavioral regularity, and inter-individual relationships. Additional tests with Japanese fire-bellied newts and Nile tilapia juveniles demonstrated comparable tracking performance in aquatic environments. Taken together, eeeHive overcomes the inherent limitations of conventional LF RFID, establishing a powerful HF RFID-based platform for fine-scale behavioral tracking of group-housed animals across terrestrial and aquatic species.

1 Introduction

The diversity and availability of model animals in neuroscience and neuropharmacology have expanded substantially over the past two decades ( 1, 2). However, the reliable interpretation of their behavior in laboratory environments remains a persistent challenge ( 3– 6). Conventional behavioral test paradigms, particularly in rodents, typically involve manually transferring animals from their home cages to isolated testing chambers for short observation periods ( 7). Since around the 2000s, there has been criticism of such “closed-session” procedures for relying on operational definitions that disregard the impact of human intervention and the sensory and behavioral traits unique to each species or strain ( 8, 9).

To address this problem, the concept of automated home-cage monitoring (AHCM) has emerged, which enables long-term and undisturbed observation of animals in their living environment, facilitating the capture of a more diverse behavioral repertoire ( 10– 14). Among AHCM approaches, RFID has become a widely adopted technology for reliable individual identification and precise behavioral tracking in group-housed environments ( 15).

RFID is a non-contact communication technology widely used for the identification and tracking of physical objects and living organisms across diverse fields, including logistics and livestock management ( [16](https://www.biorxiv.org/content/10.64898/2026.04.30.720993v

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Key points for the wiki

  • eeeHive = high-frequency (HF) RFID tracking with custom hardware: high-speed multiplexed antenna polling (5.9 ms per antenna) + concurrent multi-tag reading (8.2 ms per tag) — overcomes LF-RFID limits of poor spatiotemporal resolution and single-tag reads.
  • Validated on 24 mice (1 week), six common marmosets (7 weeks), plus Japanese fire-bellied newts and Nile tilapia juveniles (aquatic); tracks individuals even in dense clusters.
  • Extracts spatial utilisation, temporal dynamics, behavioural regularity, inter-individual relationships.
  • Lineage: complements the TraffiCage (NewBehavior AG) tile-antenna and IntelliCage ring-antenna approaches; commercialised by Phenovance LLC (Japan); the eeeHive 2D product is referenced by the IntelliProfiler 2.0 home-cage analysis pipeline (2026).

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