Abstract
The cell nucleus contains different domains and nuclear bodies, whose position relative to each other inside the nucleus can vary depending on the physiological state of the cell. Changes in the three-dimensional organization are associated with the mobility of individual components of the nucleus. In this chapter, we present a protocol for live-cell imaging and analysis of nuclear body mobility. Unlike other similar protocols, our image analysis pipeline includes non-rigid compensation for global motion of the nucleus before particle tracking and trajectory analysis, leading to precise detection of intranuclear movements. The protocol described can be easily adapted to work with most cell lines and nuclear bodies.
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Acknowledgments
We are grateful to Dr. X.W. Wang for the GFP-NPM WT-expressing plasmid. This work was supported by the Russian Science Foundation (17-11-01279 to DVS for image analysis) and the Russian Foundation for Basic Research (project 18-54-16002 to EVS for cell preparation and microscopy).
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Sorokin, D.V., Arifulin, E.A., Vassetzky, Y.S., Sheval, E.V. (2020). Live-Cell Imaging and Analysis of Nuclear Body Mobility. In: Hancock, R. (eds) The Nucleus . Methods in Molecular Biology, vol 2175. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0763-3_1
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DOI: https://doi.org/10.1007/978-1-0716-0763-3_1
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