title: Monitoring of chromosome dynamics of single yeast cells in a microfluidic platform with aperture cell traps.
abstract: Chromosome movement plays important roles in DNA replication, repair, genetic recombination, and epigenetic phenomena during mitosis and meiosis. In particular, chromosome movement in the nuclear space is essential for the reorganization of the nucleus. However, conventional methods for analyzing the chromosome movements in vivo have been limited by technical constraints of cell trapping, cell cultivation, oxygenation, and in situ imaging. Here, we present a simple microfluidic platform with aperture-based cell trapping arrays to monitor the chromosome dynamics in single living cells for a desired period of time. Under the optimized conditions, our microfluidic platform shows a single-cell trapping efficiency of 57%. This microfluidic approach enables in situ imaging of intracellular dynamics in living cells responding to variable input stimuli under the well-controlled microenvironment. As a validation of this microfluidic platform, we investigate the fundamental features of the dynamic cellular response of the individual cells treated with different stimuli and drug. We prove the basis for dynamic chromosome movement in single yeast cells to be the telomere and nuclear envelope ensembles that attach to and move in concert with nuclear actin cables. Therefore, these results illustrate the monitoring of cellular functions and obtaining of dynamic information at a high spatiotemporal resolution through the integration of a simple microfluidic platform.
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Si Hyung
authors.initials: SH
authors.lastname: Jin
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Sung-Chan
authors.initials: SC
authors.lastname: Jang
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Byungjin
authors.initials: B
authors.lastname: Lee
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Heon-Ho
authors.initials: HH
authors.lastname: Jeong
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Seong-Geun
authors.initials: SG
authors.lastname: Jeong
authors.affiliation: Institute of Biochemistry, ETH Zürich, Zürich, CH 8093, Switzerland. leesu@ ethz.ch and Scientific Center for Optical and Electron Microscopy (ScopeM), ETH Zürich, Zürich, CH-8093, Switzerland.
authors.forename: Sung Sik
authors.initials: SS
authors.lastname: Lee
authors.affiliation: Department of Life Science, Chung-Ang University, Seoul 156-756, Republic of Korea.
authors.forename: Keun Pil
authors.initials: KP
authors.lastname: Kim
authors.affiliation: Department of Chemical Engineering, Chungnam National University, 99 Daehak-ro, Yuseong-Gu, Daejeon 305-764, Republic of Korea. rhadum@cnu.ac.kr.
authors.forename: Chang-Soo
authors.initials: CS
authors.lastname: Lee
mesh_terms: D015825:Chromosomes, Fungal / Q000378:metabolism*; D004867:Equipment Design; D056656:Lab-On-A-Chip Devices; D008540:Meiosis; D059010:Single-Cell Analysis / Q000295:instrumentation*; D015003:Yeasts / Q000166:cytology* / Q000235:genetics