Microsystems for cell-based electrophysiology

Among the electrophysiology techniques, the voltage clamp and its subsequent scaling to smaller mammalian cells, the so-called patch clamp, led to fundamental discoveries in the last century, revealing the ionic mechanisms and the role of single-ion channels in the generation and propagation of action potentials through excitable membranes (e. g. nerves and muscles). Since then, these techniques have gained a reputation as the gold standard of studying cellular ion channels owing to their high accuracy and rich information content via direct measurements under a controlled membrane potential. However, their delicate and skill-laden procedure has put a serious constrain on the throughput and their immediate utilization in the discovery of new cures targeting ion channels until researchers discovered 'lab-on-a-chip' as a viable platform for the automation of these techniques into a reliable high-throughput screening functional assay on ion channels. This review examines the innovative 'lab-on-a-chip' microtechnologies demonstrated towards this target over a period of slightly more than a decade. The technologies are categorically reviewed according to their considerations for design, fabrication, as well as microfluidic integration from a performance perspective with reference to their ability to secure G Omega seals (gigaseals) on cells, the norm broadly accepted among electrophysiologists for quality recordings that reflect ion-channel activity with high fidelity

Label-free electrical quantification of amplified nucleic acids through nanofluidic diodes

A label-free method of quantifying nucleic acids in polymerase chain reaction (PCR) is described and could be the basis for miniaturized devices that can amplify and detect target nucleic acids in real time. The method takes advantage of ionic current rectification effect discovered in nanofluidic channels exhibiting a broken symmetry in electrochemical potential - nanofluidic diodes. Nanofluidic diodes are prototyped here on nanopipettes readily pulled from individual thin-walled glass capillaries for a proof of concept demonstration yet the basic concept would be applicable to ionic rectifiers constructed through other means. When a nanopipette modified in the tip region with cationic polyelectrolytes is presented with an unpurified PCR product, the tip surface electrostatically interacts with the amplicons and modulates its ionic rectification direction in response to the intrinsic charge of those adsorbed. Modulations are gradual and correlate well with the mass concentration of the amplicons above 2.5 ng/mu L, rather than their sizes, with adequate discrimination against the background. Moreover, the tip surface, following a measurement, is regenerated through a layer-by-layer assembly of cationic polyelectrolytes and amplicons. The regenerated tips are capable of measuring distinct mass concentrations without signs of noticeable degradation in sensitivity. Further, the tips are shown capable of reproducing the amplification curve of real-time PCR through sequential steps of surface regeneration and simple electrical readout during the intermediate reaction stages. This suggests that nanopipettes as nanofluidic diodes are at a capacity to be employed for monitoring the PCR progress. (C) 2013 Elsevier B.V. All rights reserved

Microchannel plate as a novel bipolar electrode for high-performance enrichment of anions

Microchannel plate (MCP), a high-porosity glass membrane used as an electron multiplier in analytical/scientific instruments for the detection of energetic photons and charged particles is demonstrated here as a highly effective bipolar electrode (BPE) for electrokinetic focusing of anions. Assembled between a pair of microfluidic channels filled with an electrolyte buffer and subjected to a sufficient bias potential, MCP supports faradaic reactions, owing to its semiconducting characteristics. Thousands of microcapillary tubes fused together define MCP and act in unison such that each microcapillary serves as a tiny BPE surrounding an infinitesimal element of bulk electrolyte with a large surface-area-to-volume ratio and hence performs highly effective as compared to a planar electrode inlaid into a microchannel. This performance has been validated here where concentration enrichment of a fluorescent tracer has been demonstrated at a remarkable rate of up to 175-fold/s exceeding those reported for planar BPEs. We attribute such high performance to the rapid onset of ion-depletion zone and subsequent steep field gradient, signifying the high-porosity structure of MCP as an effective BPE

Cylindrical glass nanocapillaries patterned via coarse lithography (>1 μm) for biomicrofluidic applications

We demonstrate a new method of fabricating in-plane cylindrical glass nanocapillaries (< 100 nm) that does not require advanced patterning techniques but the standard coarse photolithography (> 1 mu m). These nanocapillaries are self-enclosed optically transparent and highly regular over large areas. Our method involves structuring mu m-scale rectangular trenches in silicon, sealing the trenches into enclosed triangular channels by depositing phosphosilicate glass, and then transforming the channels into cylindrical capillaries through shape transformation by the reflow of annealed glass layer. Extended anneal has the structures shrunk into nanocapillaries preserving their cylindrical shape. Nanocapillaries similar to 50 nm in diameter and effective stretching of digested lambda-phage DNA in them are demonstrated. (C) 2012 American Institute of Physics. [http://dx.doi.org/10.1063/1.4771691

Induced hydraulic pumping via integrated submicrometer cylindrical glass capillaries

Here, we report on a micropump that generates hydraulic pressure owing to a mismatch in EOF rates of microchannels and submicrometer cylindrical glass capillaries integrated on silicon. The electrical conductance of such capillaries in the dilute limit departs from bulk linear behavior as well as from the surface-charge-governed saturation in nanoslits that is well described by the assumption of a constant surface charge density. The capillaries show rather a gradual decrease in conduction at low salt concentrations, which can be explained more aptly by a variable surface charge density that accounts for chemical equilibrium of the surface. The micropump uses a traditional cross-junction structure with ten identical capillaries integrated in parallel on a side arm and each with a 750 nm diameter and 3 mm length. For an applied voltage of 700 V, a hydraulic pressure up to 5 kPa is generated with a corresponding flow velocity nearly 3 mm/s in a straight field-free branch 20 mu m wide, 10 mu m deep, and 10 mm long. The micropump utility has been demonstrated in an open tubular LC of three fluorescently labeled amino acids in just less than 20 s with minimal plate height values between 3 and 7 mu m. The submicrometer capillaries are self-enclosed and produced through a unique process that does not require high-resolution advanced lithography or wafer-bonding techniques to define their highly controlled precise structures