Affiliation:
1. i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal;
2. Departamento de Biomedicina, Faculdade de Medicina, Universidade do Porto, Portugal.
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 to track centrosomes and reconstruct 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/analysis algorithms. It provides detailed quantifications of the spatiotemporal relations between centrosomes, nucleus 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 with the mechanical forces exerted on the nucleus by its adjacent cytoskeletal structures. Unlike previous algorithms based on circular/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 to help unravel new elements in the spatiotemporal dynamics of subcellular structures.
Funder
Fundação para a Ciência e a Tecnologia
Publisher
The Company of Biologists
Cited by
3 articles.
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