Kinetochore tracking in 3D from lattice light-sheet imaging data with KiT.

Kinetochore tracking in 3D from lattice light-sheet imaging data with KiT.
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DOI:
10.1093/bioinformatics/btac330
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发表时间:
2022-06-13
期刊:
Bioinformatics (Oxford, England)
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晶格光片显微镜(LLSM)正在彻底改变细胞生物学,因为它能够实现快速,高分辨率的三维扩展成像,并大大减少光毒性和漂白。然而,对这些数据集的分析仍然是一个重大挑战。着丝粒是促进和控制有丝分裂纺锤体内染色体微管附着的蛋白质复合物,其自动跟踪提供了有丝分裂中染色体动力学的定量评估。在这里,我们扩展现有的开源着丝粒跟踪软件(KiT)跟踪(和配对)在LLSM数据中的整个前中期到后期的着丝粒。其中一个关键的改进是目标函数中的正则化项,以强制执行有关人类有丝分裂细胞中动粒数量的生物信息,以及改进的诊断工具。该软件提供了关于着丝粒如何在有丝分裂期间稳健地确保染色体的聚集和分离的定量见解。KiT是在MATLAB中实现的免费开源软件,可以从https://github.com/cmcb-warwick/KiT下载。源代码库可在https://bitbucket.org/jarmond/kit(标签v2.4.0)上获得,并正在继续开发中。 补充数据可在Bioinformatics在线获得。
Lattice light-sheet microscopy (LLSM) is revolutionizing cell biology since it enables fast, high-resolution extended imaging in three dimensions combined with a drastic reduction in photo-toxicity and bleaching. However, analysis of such datasets still remains a major challenge. Automated tracking of kinetochores, the protein complex facilitating and controlling microtubule attachment of the chromosomes within the mitotic spindle, provides quantitative assessment of chromosome dynamics in mitosis. Here, we extend existing open-source kinetochore tracking software (KiT) to track (and pair) kinetochores throughout prometaphase to anaphase in LLSM data. One of the key improvements is a regularization term in the objective function to enforce biological information about the number of kinetochores in a human mitotic cell, as well as improved diagnostic tools. This software provides quantitative insights into how kinetochores robustly ensure congression and segregation of chromosomes during mitosis. KiT is free, open-source software implemented in MATLAB and can be downloaded as a package from https://github.com/cmcb-warwick/KiT. The source repository is available at https://bitbucket.org/jarmond/kit (tag v2.4.0) and under continuing development. Supplementary data are available at Bioinformatics online.
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期刊: JOURNAL OF MICROSCOPY-OXFORD
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发表时间: 2014-10-24
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