Waipot Ngamsaad

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

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.

Theoretical studies of phase-separation kinetics in a Brinkman porous medium

Although the coarsening of binary fluid mixtures in porous media has been of great interest for some time, there are still no complete theories for describing the relevant mechanisms, and more theoretical work needs to be carried out. In this work, we have proposed a simple model for phase separation of binary fluids in a porous medium, where the Brinkman-extended-Darcy equation and Cahn–Hilliard equation are the dynamical constitutions. Using the dimensional analysis approach, our findings lead to the prediction of domain coarsening in a porous medium for several regimes, including the conventional power laws. In addition, we have found that slowed-down coarsening dynamics are caused by the hydrodynamic screening effect, which is governed by the logarithmic law for this regime. Our theoretical results are at least qualitatively consistent with previous reports using simulations or experiments.

Pinning of domains for fluid–fluid phase separation in lipid bilayers with asymmetric dynamics

We propose a physical mechanism for the arrest of domain coarsening in a system of two apposed two-dimensional binary fluids. The two fluids are subject to a dynamic asymmetry: strong friction with the environment allows domains in one fluid layer (the “bottom” fluid) to grow only diffusively, whereas hydrodynamic flow leads to initially faster growth in the apposed fluid (the “top” layer). The two fluids are energetically coupled so that domains of similar type interact favorably across the two fluids. Using lattice Boltzmann simulations we observe that at a certain length scale, which is independent of the coarsening state in the bottom layer, domain growth in the top layer comes to an arrest. A phenomenological model suggests the pinning of domains across the two fluids to cause the arrest in domain growth. The pinning results from the interplay between line tension and domain coupling strength across the two fluids. We apply our model to a lipid bilayer for which we calculate the length scale of the dynamically arrested domains in the top layer. We find domain extensions of about or somewhat larger than 20 nm. Potential applications of our pinning model are to mixed lipid bilayers that tend to phase separate and are subject to a dynamic asymmetry; these include model membranes on a solid support and lipid rafts in the plasma membrane.