Quantitative analysis of time-series fluorescence microscopy using a spot tracking method: application to Min protein dynamics |
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Authors: | Somrit Unai Paisan Kanthang Udorn Junthon Waipot Ngamsaad Wannapong Triampo Charin Modchang Chartchai Krittanai |
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Institution: | (1) R&D Group of Biological and Environmental Physics, Department of Physics, Faculty of Science, Mahidol University, Rama 6 Rd., Ratchatewee, Bangkok, 10400, Thailand;(2) Center of Excellence for Vector and Vector-Borne Diseases, Faculty of Science, Mahidol University, Nakhonpathom, Thailand;(3) Institute for Innovation and Development of Learning Process, Mahidol University, Bangkok, Thailand;(4) Institute of Molecular Biology. and Genetics, Mahidol University, Nakhonpathom, Thailand |
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Abstract: | The dynamics of MinD protein has been recognized as playing an important role in the accurate positioning of the septum during
cell division. In this work, spot tracking technique (STT) was applied to track the motion and quantitatively characterize
the dynamic behavior of green fluorescent protein-labeled MinD (GFP-MinD) in an Escherichia coli system. We investigated MinD dynamics on the level of particle ensemble or cluster focusing on the position and motion of
the maximum in the spatial distribution of MinD proteins. The main results are twofold: (i) a demonstration of how STT could
be an acceptable tool for MinD dynamics studies; and (ii) quantitative findings with parametric and non-parametric analyses.
Specifically, experimental data monitored from the dividing E. coli cells (typically 4.98 ± 0.75 μm in length) has demonstrated a fast oscillation of the MinD protein between the two poles,
with an average period of 54.6 ± 8.6 s. Observations of the oscillating trajectory and velocity show a trapping or localized
behavior of MinD around the polar zone, with average localization velocity of 0.29 ± 0.06 μm/s; and flight switching was observed
at the pole-to-pole leading edge, with an average switching velocity of 2.95 ± 0.31 μm/s. Subdiffusive motion of MinD proteins
at the polar zone was found and investigated with the dynamic exponent, α of 0.34 ± 0.16. To compare with the Gaussian-based analysis, non-parametric statistical analysis and noise consideration
were also performed.
Co-first authors |
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Keywords: | spot tracking E coli cell division Min proteins MinD protein oscillation |
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