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1.
Mol Biol Cell ; 28(12): 1601-1611, 2017 Jun 15.
Article in English | MEDLINE | ID: mdl-28450455

ABSTRACT

Faithful segregation of chromosomes during cell division relies on multiple processes such as chromosome attachment and correct spindle positioning. Yet mitotic progression is defined by multiple parameters, which need to be quantitatively evaluated. To study the spatiotemporal control of mitotic progression, we developed a high-content analysis (HCA) approach that combines automated fluorescence microscopy with real-time quantitative image analysis and allows the unbiased acquisition of multiparametric data at the single-cell level for hundreds of cells simultaneously. The Mitotic Analysis and Recording System (MAARS) provides automatic and quantitative single-cell analysis of mitotic progression on an open-source platform. It can be used to analyze specific characteristics such as cell shape, cell size, metaphase/anaphase delays, and mitotic abnormalities including spindle mispositioning, spindle elongation defects, and chromosome segregation defects. Using this HCA approach, we were able to visualize rare and unexpected events of error correction during anaphase in wild-type or mutant cells. Our study illustrates that such an expert system of mitotic progression is able to highlight the complexity of the mechanisms required to prevent chromosome loss during cell division.


Subject(s)
Chromosome Segregation/physiology , Image Processing, Computer-Assisted/methods , Single-Cell Analysis/methods , Cell Cycle Proteins/genetics , Chromosome Segregation/genetics , Kinetochores/physiology , Mitosis/genetics , Mitosis/physiology , Saccharomycetales/genetics , Schizosaccharomyces/genetics , Software , Spatio-Temporal Analysis , Spindle Apparatus/physiology
2.
J Cell Biol ; 212(7): 747-9, 2016 Mar 28.
Article in English | MEDLINE | ID: mdl-27002164

ABSTRACT

Increasing evidence in eukaryotic cells suggests that mechanical forces are essential for building a robust mitotic apparatus and correcting inappropriate chromosome attachments. In this issue, Cojoc et al. (2016. J. Cell Biol., http://dx.doi.org/10.1083/jcb.201506011) use laser microsurgery in vivo to measure and study the viscoelastic properties of kinetochores.


Subject(s)
Chromosome Segregation , Kinetochores/metabolism , Laser Therapy , Mechanotransduction, Cellular , Microsurgery , Microtubules/physiology , Schizosaccharomyces/physiology , Humans
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