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Open AccessArticle

Mitotic Chromosomes in Live Cells Characterized Using High-Speed and Label-Free Optical Diffraction Tomography

1
Asan Institute for Life Sciences, Asan Medical Center, Seoul 05505, Korea
2
Division of Physical Pharmacy, Faculty of Pharmaceutical Sciences, Kobe Gakuin University, Kobe 650-8586, Japan
3
Tomocube Inc., Daejeon 34051, Korea
4
Department of Physics, Korea Advanced Institute of Science and Technology, Daejeon 34141, Korea
5
Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul 05505, Korea
6
Department of Radiation Oncology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, Korea
*
Authors to whom correspondence should be addressed.
These authors contributed equally to this work.
Cells 2019, 8(11), 1368; https://doi.org/10.3390/cells8111368
Received: 18 September 2019 / Revised: 21 October 2019 / Accepted: 30 October 2019 / Published: 31 October 2019
(This article belongs to the Special Issue Nuclear Organisation)
The cell nucleus is a three-dimensional, dynamic organelle organized into subnuclear compartments such as chromatin and nucleoli. The structure and function of these compartments are maintained by diffusion and interactions between related factors as well as by dynamic and structural changes. Recent studies using fluorescent microscopic techniques suggest that protein factors can access and are freely mobile in heterochromatin and in mitotic chromosomes, despite their densely packed structure. However, the physicochemical properties of the chromosome during cell division are not fully understood. In the present study, characteristic properties such as the refractive index (RI), volume of the mitotic chromosomes, and diffusion coefficient (D) of fluorescent probes inside the chromosome were quantified using an approach combining label-free optical diffraction tomography with complementary confocal laser-scanning microscopy and fluorescence correlation spectroscopy. Variations in these parameters correlated with osmotic conditions, suggesting that changes in RI are consistent with those of the diffusion coefficient for mitotic chromosomes and cytosol. Serial RI tomography images of chromosomes in live cells during mitosis were compared with three-dimensional confocal micrographs to demonstrate that compaction and decompaction of chromosomes induced by osmotic change were characterized by linked changes in chromosome RI, volume, and the mobilities of fluorescent proteins. View Full-Text
Keywords: optical diffraction tomography; fluorescence correlation spectroscopy; mitosis; chromosome; refractive index; cellular viscosity; diffusion coefficient; osmotic stress optical diffraction tomography; fluorescence correlation spectroscopy; mitosis; chromosome; refractive index; cellular viscosity; diffusion coefficient; osmotic stress
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Kim, T.-K.; Lee, B.-W.; Fujii, F.; Lee, K.-H.; Lee, S.; Park, Y.; Kim, J.K.; Lee, S.-W.; Pack, C.-G. Mitotic Chromosomes in Live Cells Characterized Using High-Speed and Label-Free Optical Diffraction Tomography. Cells 2019, 8, 1368.

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