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Analyzing chromosome condensation in yeast by second-harmonic generation microscopy

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Abstract

Condensation is a fundamental property of mitotic chromosomes in eukaryotic cells. However, analyzing chromosome condensation in yeast is a challenging task while existing methods have notable weaknesses. Second-harmonic generation (SHG) microscopy is a label-free, advanced imaging technique for measuring the surface curve of isotropic molecules such as chromatin in live cells. We applied this method to detect changes in chromatin organization throughout the cell cycle in live yeast cells. We showed that SHG microscopy can be used to identify changes in chromatin organization throughout the cell cycle and in response to inactivation of the SMC complexes, cohesin and condensin. Implementation of this method will improve our ability to analyze chromatin structure in protozoa and will enhance our understanding of chromatin organization in eukaryotic cells.

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Acknowledgements

We thank Jennifer Gerton and Maya Schuldiner for providing yeast strains. Eyla Maoz and Yehuda Gabay for their help with cell cytometry. We also thank members of the Onn lab for their support.

Funding

This work was supported by the Israel Science Foundation Grant 1099/16 (IO)

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Authors and Affiliations

  1. Chromosome Instability and Dynamics Laboratory, The Azrieli Faculty of Medicine, Bar-Ilan University, Safed, Israel

    Katreena Yamin, Avi Matityahu & Itay Onn

  2. Imaging Unit, The Azrieli Faculty of Medicine, Bar-Ilan University, Safed, Israel

    Michael Assa

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  1. Katreena Yamin
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  2. Michael Assa
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  3. Avi Matityahu
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  4. Itay Onn
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Correspondence to Itay Onn.

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Communicated by M. Kupiec.

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Yamin, K., Assa, M., Matityahu, A. et al. Analyzing chromosome condensation in yeast by second-harmonic generation microscopy. Curr Genet 66, 437–443 (2020). https://doi.org/10.1007/s00294-019-01034-1

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  • DOI: https://doi.org/10.1007/s00294-019-01034-1

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