Journal of neuroscience methods · 2026
A centroid-based video-tracking framework for integrated analysis of sleep, locomotor activity, and spatial behavior in Drosophila melanogaster.
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Duarte Tâmie, Blache Dominique, Corte Cristiane Lenz Dalla
Аннотация
BACKGROUND: Sleep in Drosophila melanogaster is commonly quantified using infrared beam-break systems, which are robust for high-throughput monitoring but provide limited information about the spatial and temporal organization of behavior. We proposed that continuous centroid-based video tracking could provide an accessible, no-code framework that simultaneously extracts sleep-related immobility, locomotor activity, temporal movement profiles, and spatial behavioral readouts.
NEW METHOD: A centroid-based video-tracking framework was developed in ANY-maze for integrated analysis of sleep, locomotor, temporal, and spatial occupancy in D. melanogaster. Individual flies were recorded in horizontal tubes, and the workflow extracted immobility-defined sleep, sleep architecture, locomotor output, activity distribution, and spatial occupancy. Analytical reproducibility was assessed through repeated analysis of identical recordings, and sleep-related outputs were compared with DAM2 measures under light-dark (LD) and constant-light (LL) conditions.
RESULTS: The framework showed high analytical reproducibility across outputs. Under LL, canonical sleep phenotypes were detected, including reduced total sleep and fragmented sleep architecture. Continuous tracking also detected changes in spatial occupancy and temporal movement organization that were not captured by cumulative activity measures.
COMPARISON WITH EXISTING METHODS: Biological outcomes for LL-induced sleep disruption were similar between ANY-maze and DAM2. We discuss methodological differences between continuous centroid tracking and event-based beam-break monitoring. Unlike beam-based systems, the ANY-maze workflow preserves spatial and temporal information without custom programming.
CONCLUSIONS: The no-code, video-based ANY-maze framework provides an accessible, reproducible strategy for multidimensional behavioral phenotyping in fruit flies and extends behavioral interpretation beyond total sleep or activity measures while remaining compatible with standard laboratory setups.
NEW METHOD: A centroid-based video-tracking framework was developed in ANY-maze for integrated analysis of sleep, locomotor, temporal, and spatial occupancy in D. melanogaster. Individual flies were recorded in horizontal tubes, and the workflow extracted immobility-defined sleep, sleep architecture, locomotor output, activity distribution, and spatial occupancy. Analytical reproducibility was assessed through repeated analysis of identical recordings, and sleep-related outputs were compared with DAM2 measures under light-dark (LD) and constant-light (LL) conditions.
RESULTS: The framework showed high analytical reproducibility across outputs. Under LL, canonical sleep phenotypes were detected, including reduced total sleep and fragmented sleep architecture. Continuous tracking also detected changes in spatial occupancy and temporal movement organization that were not captured by cumulative activity measures.
COMPARISON WITH EXISTING METHODS: Biological outcomes for LL-induced sleep disruption were similar between ANY-maze and DAM2. We discuss methodological differences between continuous centroid tracking and event-based beam-break monitoring. Unlike beam-based systems, the ANY-maze workflow preserves spatial and temporal information without custom programming.
CONCLUSIONS: The no-code, video-based ANY-maze framework provides an accessible, reproducible strategy for multidimensional behavioral phenotyping in fruit flies and extends behavioral interpretation beyond total sleep or activity measures while remaining compatible with standard laboratory setups.