An efficient kinetic Monte Carlo to study analyte capture by a nanopore: transients, boundary conditions and time-dependent fields.

To better understand the capture process by a nanopore, we introduce an efficient Kinetic Monte Carlo (KMC) algorithm that can simulate long times and large system sizes by mapping the dynamic of a point-like particle in a 3D spherically symmetric system onto the 1D biased random walk. Our algorithm recovers the steady-state analytical solution and allows us to study time-dependent processes such as transients. Simulation results show that the steady-state depletion zone near pore is barely larger than the pore radius and narrows at higher field intensities; as a result, the time to reach steady-state is much smaller than the time required to empty a zone of the size of the capture radius λe. When the sample reservoir has a finite size, a second depletion region propagates inward from the outer wall, and the capture rate starts decreasing when it reaches the capture radius λe. We also note that the flatness of the electric field near the pore, which is often neglected, induces a traffic jam that can increase the transient time by several orders of magnitude. Finally, we propose a new proof-of-concept scheme to separate two analytes of the same mobility but different diffusion coefficients using time-varying fields.

Voltage-driven translocation: Defining a capture radius.

Analyte translocation involves three phases: (i) diffusion in the loading solution, (ii) capture by the pore, and (iii) threading. The capture process remains poorly characterized because it cannot easily be visualized or inferred from indirect measurements. The capture performance of a device is often described by a capture radius generally defined as the radial distance R* at which diffusion-dominated dynamics cross over to field-induced drift. However, this definition is rather ambiguous and the related models are usually oversimplified and studied in the steady-state limit. We investigate different approaches to defining and estimating R* for a charged particle diffusing in a liquid and attracted to the nanopore by the electric field. We present a theoretical analysis of the Péclet number as well as Monte Carlo simulations with different simulation protocols. Our analysis shows that the boundary conditions, pore size, and finite experimental times all matter in the interpretation and calculation of R*.

Physical confinement signals regulate the organization of stem cells in three dimensions.

During embryogenesis, the spherical inner cell mass (ICM) proliferates in the confined environment of a blastocyst. Embryonic stem cells (ESCs) are derived from the ICM, and mimicking embryogenesis in vitro, mouse ESCs (mESCs) are often cultured in hanging droplets. This promotes the formation of a spheroid as the cells sediment and aggregate owing to increased physical confinement and cell-cell interactions. In contrast, mESCs form two-dimensional monolayers on flat substrates and it remains unclear if the difference in organization is owing to a lack of physical confinement or increased cell-substrate versus cell-cell interactions. Employing microfabricated substrates, we demonstrate that a single geometric degree of physical confinement on a surface can also initiate spherogenesis. Experiment and computation reveal that a balance between cell-cell and cell-substrate interactions finely controls the morphology and organization of mESC aggregates. Physical confinement is thus an important regulatory cue in the three-dimensional organization and morphogenesis of developing cells.