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Automated Optimization of Bacterial Tracking Pipelines with TrackMate 8
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Paper
Automated Optimization of Bacterial Tracking Pipelines with TrackMate 8
2026
Overview
Quantitative analysis of bacterial dynamics in time-lapse microscopy requires robust tracking pipelines, yet selecting and optimizing algorithms for specific experiments remains challenging. Indeed, Microbiologists are confronted with numerous algorithms that must be carefully chosen and parameterized to achieve optimal tracking for their experiments. We present an automated methodology to determine optimal tracking configurations for microbiological applications. It is based on TrackMate 8, a novel version of the TrackMate Fiji plugin extended with microbiology-specific tools. Our approach systematically evaluates algorithm-parameter combinations optimizing biologically relevant metrics (e.g., cell-cycle accuracy, bacteria morphology) and includes: (1) integration of deep-learning algorithms (Omnipose, YOLO, Trackastra) adequate for bacteria images in TrackMate, (2) a TrackMate-Helper extension for parameter optimization, and (3) a tracking and segmentation editor for tracking ground-truth generation. We demonstrate the effectiveness of the methodology on two use cases showing its adaptability to diverse experimental conditions. This methodology enables microbiologists with a widely applicable, automated framework to optimize tracking pipelines, facilitating quantitative analysis in bacterial imaging.Competing Interest StatementThe authors have declared no competing interest.Footnotes* Fixed upload of badly formatted Supplemental Information file.* https://zenodo.org/records/17909896* https://zenodo.org/records/17911259Funder Information DeclaredAgence Nationale de la Recherche, ANR-24-INBS-0005 FBI BIOGEN, ANR-10-PATH-003 HELDIVPAT, ANR-10-LBX-62 IBEID, ANR-16-CONV-0005 INCEPTION, ANR-17-EURE-0012 EURIP, ANR-19-CE44-0014O2-TABOOEuropean Research Council, DESTOP European Research Council (ERC) Advanced grant (101097791), PGNfromSHAPEtoVIR, FP7-202283, IMI 2 Joint Undertaking (JU) under Grant Agreement No 853989Fondation pour la Recherche Médicale, EQU202403018034, FDT202504020138Gates Foundation, IMI 2 Joint Undertaking (JU) under Grant Agreement No 853989
Publisher
Cold Spring Harbor Laboratory Press
Subject
