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Methods Cell Biol. 2018;145:1-27. doi: 10.1016/bs.mcb.2018.03.009.

Mitotic live-cell imaging at different timescales.

Author information

1
Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland.
2
Centre for Mechanochemical Cell Biology & Division of Biomedical Science, Warwick Medical School, University of Warwick, Coventry, United Kingdom.
3
Mathematics Institute, University of Warwick, Coventry, United Kingdom.
4
Centre for Mechanochemical Cell Biology & Division of Biomedical Science, Warwick Medical School, University of Warwick, Coventry, United Kingdom. Electronic address: a.d.mcainsh@warwick.ac.uk.
5
Department of Cell Physiology and Metabolism, University of Geneva, Geneva, Switzerland. Electronic address: patrick.meraldi@unige.ch.

Abstract

Mitosis is a highly dynamic and choreographed process in which chromosomes are captured by the mitotic spindle and physically segregated into the two daughter cells to ensure faithful transmission of the genetic material. Live-cell fluorescence microscopy enables these dynamics to be analyzed over diverse temporal scales. Here we present the methodologies to study chromosome segregation at three timescales: we first show how automated tracking of kinetochores enables investigation of mitotic spindle and chromosome dynamics in the seconds-to-minutes timescale; next we highlight how new DNA live dyes allow the study of chromosome segregation over a period of several hours in any cell line; finally, we demonstrate how image sequences acquired over several days can reveal the fate of whole cell populations over several consecutive cell divisions.

KEYWORDS:

Chromosome segregation; Chromosome segregation error; Kinetochore; Live-cell imaging; Microtubule dynamics; Mitosis

PMID:
29957199
DOI:
10.1016/bs.mcb.2018.03.009
[Indexed for MEDLINE]

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