Rheology and simulation of 2-dimensional clathrin protein network assembly.

Clathrin is a three-legged protein complex that assembles into lattice structures on the cell membrane and transforms into fullerene-like cages during endocytosis. This dynamic structural flexibility makes clathrin an attractive building block for guided assembly. The assembly dynamics and the mechanical properties of clathrin protein lattices are studied using rheological measurements and theoretical modelling in an effort to better understand two dynamic processes: protein adsorption to the interface and assembly into a network. We find that percolation models for protein network formation are insufficient to describe clathrin network formation, but with Monte Carlo simulations we can describe the dynamics of network formation very well. Insights from this work can be used to design new bio-inspired nano-assembly systems.

Nanostraws for direct fluidic intracellular access.

Nanomaterials are promising candidates to improve the delivery efficiency and control of active agents such as DNA or drugs directly into cells. Here we demonstrate cell-culture platforms of nanotemplated "nanostraws" that pierce the cell membrane, providing a permanent fluidic pipeline into the cell for direct cytosolic access. Conventional polymeric track-etch cell culture membranes are alumina coated and etched to produce fields of nanostraws with controllable diameter, thickness, and height. Small molecules and ions were successfully transported into the cytosol with 40 and 70% efficiency, respectively, while GFP plasmids were successfully delivered and expressed. These platforms open the way for active, reproducible delivery of a wide variety of species into cells without endocytosis.

Biomimetic surface patterning for long-term transmembrane access.

Here we present a planar patch clamp chip based on biomimetic cell membrane fusion. This architecture uses nanometer length-scale surface patterning to replicate the structure and function of membrane proteins, creating a gigaohm seal between the cell and a planar electrode array. The seal is generated passively during cell spreading, without the application of a vacuum to the cell surface. This interface can enable cell-attached and whole-cell recordings that are stable to 72 hours, and generates no visible damage to the cell. The electrodes can be very small (<5 µm) and closely packed, offering a high density platform for cellular measurement.

A microfluidic approach to synthesizing high-performance microfibers with tunable anhydrous proton conductivity.

Here, we demonstrate a new approach for the synthesis of ion exchange microfibers with finely tuned anhydrous conductivity. This work presents microfluidics as a system to control the size and phosphoric acid (PA) doping level of the polybenzimidazole (PBI) microfibers. It has been shown that the PA doping level can be controlled by varying the flow ratios in the microfluidic channel. The diameter of the microfibers increased with extending mixing time, whereas the doping level decreased with increasing flow ratio. The highest doping level, 16, was achieved at the flow ratio of 0.175. The anhydrous proton conductivity of the microfibers was found to be adjustable between 0.01 and 0.1 S cm(-1) at 160 °C, which is considerably higher than for conventionally doped PBI cast membranes (0.004 S cm(-1)). Furthermore, molecular dynamic simulation of proton conduction through the microfibers at different doping levels was in good agreement with the experimental results. These results demonstrate the potential of the microfluidic technique to precisely tune the physicochemical properties of PBI microfibers for various electrochemical applications such as hydrogen sensors, fuel cells as well as artificial muscles.

Rapid spatial and temporal controlled signal delivery over large cell culture areas.

Controlled chemical delivery in microfluidic cell culture devices often relies on slowly evolving diffusive gradients, as the spatial and temporal control provided by fluid flow results in significant cell-perturbation. In this paper we introduce a microfluidic device architecture that allows for rapid spatial and temporal soluble signal delivery over large cell culture areas without fluid flow over the cells. In these devices the cell culture well is divided from a microfluidic channel located directly underneath the chamber by a nanoporous membrane. This configuration requires chemical signals in the microchannel to only diffuse through the thin membrane into large cell culture area, rather than diffuse in from the sides. The spatial chemical pattern within the microfluidic channel was rapidly transferred to the cell culture area with good fidelity through diffusion. The cellular temporal response to a step-function signal showed that dye reached the cell culture surface within 45 s, and achieved a static concentration in under 6 min. Chemical pulses of less than one minute were possible by temporally alternating the signal within the microfluidic channel, enabling rapid flow-free chemical microenvironment control for large cell culture areas.