Handling Editor: David Stephens
ABSTRACT
During the initial stages of mitosis, multiple mechanisms drive centrosome separation and positioning. How they are coordinated to promote centrosome migration to opposite sides of the nucleus remains unclear. Here, we present Trackosome, an open-source image analysis software for tracking centrosomes and reconstructing nuclear and cellular membranes, based on volumetric live-imaging data. The toolbox runs in MATLAB and provides a graphical user interface for easy access to the tracking and analysis algorithms. It provides detailed quantification of the spatiotemporal relationships between centrosomes, nuclear envelope and cellular membrane, and can also be used to measure the dynamic fluctuations of the nuclear envelope. These fluctuations are important because they are related to the mechanical forces exerted on the nucleus by its adjacent cytoskeletal structures. Unlike previous algorithms based on circular or elliptical approximations, Trackosome measures membrane movement in a model-free condition, making it viable for irregularly shaped nuclei. Using Trackosome, we demonstrate significant correlations between the movements of the centrosomes, and identify specific oscillation modes of the nuclear envelope. Overall, Trackosome is a powerful tool that can be used to help unravel new elements in the spatiotemporal dynamics of subcellular structures.
Footnotes
Competing interests
The authors declare no competing or financial interests.
Author contributions
Conceptualization: D.C., J.G.F., P.A.; Methodology: D.C., J.G.F., P.A.; Software: D.C., P.A.; Formal analysis: D.C., J.G.F., P.A.; Investigation: D.C., V.N., J.T.L., J.G.F., P.A.; Resources: J.G.F., P.A.; Data curation: D.C., V.N., J.T.L., J.G.F., P.A.; Writing - original draft: D.C., J.G.F., P.A.; Writing - review & editing: D.C., J.G.F., P.A.; Supervision: J.G.F., P.A.; Project administration: P.A.; Funding acquisition: J.G.F., P.A.
Funding
This work was funded by grants from Fundo Europeu de Desenvolvimento Regional (FEDER) through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior (FCT) in the framework of the project PTDC/BEX-BCM/1758/2014 (POCI-01–0145-FEDER-016589). V.N. is supported by grant PD/BD/135545/2018 from the BiotechHealth Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior PhD program. J.T.L. is supported by grant SFRH/BD/147169/2019 from Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior. The laboratory of Helder Maiato has received funding for research and equipment from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (grant agreement No 681443).
Supplementary information
Supplementary information available online at https://jcs.biologists.org/lookup/doi/10.1242/jcs.252254.supplemental
- Received July 26, 2020.
- Accepted November 2, 2020.
- © 2020. Published by The Company of Biologists Ltd
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