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FOCUS-3D: Robust, generalizable volumetric cell segmentation for three-dimensional fluorescence microscopy

Zhang, Q.; Mu, Z.; Liu, B.; Chi, Y.; Li, D.; Wang, W.; Ni, J.-Q.; Wan, Y.; Yu, L.; Navajas Acedo, J.; Yu, G.

2026-08-28 bioinformatics
10.64898/2026.08.25.746907 bioRxiv
Show abstract

Understanding how cells establish spatial organization within tissues is a fundamental question in life sciences. While modern three-dimensional fluorescence microscopy captures large-volume tissue architecture, extracting quantitative cellular insights from complex volumetric datasets remains a major barrier. Here, we introduce FOCUS-3D, a robust, broadly generalizable volumetric cell segmentation framework built on a large, diverse manually annotated cell resource and advanced AI designs. Integrating volumetric representation learning, multi-scale feature extraction, and query-based mask prediction, FOCUS-3D achieves state-of-the-art performance across diverse species, tissues, fluorescent reporters and imaging modalities. During zebrafish (Danio rerio) development, FOCUS-3D uncovers three successive phases of notochord morphogenesis. We disentangle early motility-driven rearrangements from later cell shape remodeling and tissue repacking, and further link these morphological states to spatial and developmental transcriptional programs across independent datasets.

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