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Quantitative analysis of time-series fluorescence microscopy using a spot tracking method: application to Min protein dynamics

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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.

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Abbreviations

GFP-MinD:

green fluorescent protein-labeled MinD protein

MTS:

membrane-targeting sequence

MSD:

mean squared displacement

PSD:

power spectrum density

ROI:

region of interest

SPT:

single particle tracking

STT:

spot tracking technique

Z-ring:

equatorial ring

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Authors and Affiliations

  1. R&D Group of Biological and Environmental Physics, Department of Physics, Faculty of Science, Mahidol University, Rama 6 Rd., Ratchatewee, Bangkok, 10400, Thailand

    Somrit Unai, Paisan Kanthang, Udorn Junthon, Waipot Ngamsaad, Wannapong Triampo & Charin Modchang

  2. Center of Excellence for Vector and Vector-Borne Diseases, Faculty of Science, Mahidol University, Nakhonpathom, Thailand

    Wannapong Triampo

  3. Institute for Innovation and Development of Learning Process, Mahidol University, Bangkok, Thailand

    Wannapong Triampo

  4. Institute of Molecular Biology. and Genetics, Mahidol University, Nakhonpathom, Thailand

    Chartchai Krittanai

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  1. Somrit Unai
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  2. Paisan Kanthang
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Correspondence to Wannapong Triampo.

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Unai, S., Kanthang, P., Junthon, U. et al. Quantitative analysis of time-series fluorescence microscopy using a spot tracking method: application to Min protein dynamics. Biologia 64, 27–42 (2009). https://doi.org/10.2478/s11756-009-0013-y

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