Free-flow biomolecular concentration and separation of proteins and nucleic acids using teíchophoresis.

The ability to preconcentrate, separate, and purify biomolecules, such as proteins and nucleic acids, is an important requirement for the next generation of portable diagnostic tools for environmental monitoring and disease detection. Traditionally, such pretreatment has been accomplished using large, centralized liquid- or solid-phase extraction equipment, which can be time-consuming and requires many processing steps. Here, we present a newly developed electrokinetic concentration technique, teíchophoresis (TPE), to concentrate and separate proteins, and to concentrate nucleic acids. In TPE, a free-flowing sample is exposed to a perpendicular electric field in the vicinity of a mass-impermeable conductive wall and a conductive terminating electrolyte (TE), which creates a high electric field strength zone between the lower mobility sample and the no-flux barrier. Unlike a similar electrokinetic concentration method, isotachophoresis (ITP), TPE does not require a leading electrolyte (LE), yet still enables a continuous field-driven electrophoretic ion migration across the channel and a free-flowing biomolecular concentration at the conductive wall. Here, we demonstrate the use of free-flow TPE (FFTPE) to manipulate biomolecular samples containing proteins or nucleic acids. We first use TPE to drive a 6.6-fold concentration increase of avidin-FITC, and also demonstrate protein separation and stacking between ovalbumin-fluorescein and BSA-AlexaFluor 555, both without the use of a conventional LE. Further, we utilize TPE to perform a 21-fold concentration increase of nucleic acids. Our results show that TPE is biocompatible with both proteins and nucleic acids, requires only 10 V DC, produces no significant sample pH changes during operation, and demonstrates that this method can be used as an effective sample pretreatment to prepare biological samples for downstream analysis in a continuous free-flowing microfluidic channel.

Continuous Molecular Concentration and Separation Using Pulsed-Field Conductive-Wall Single-Buffer Teíchophoresis.

We present an experimental study of a novel continuous electrokinetic molecular concentration and separation technique termed teíchophoresis (TPE). We demonstrate here that TPE can serve as a potential alternative to the electrokinetic method isotachophoresis (ITP). In ITP, an electric field serves to focus charged species between a low-mobility terminating electrolyte (TE) and a high-mobility leading electrolyte (LE). Similarly, TPE serves to focus charged species between a low-mobility TE; however, the LE is conveniently replaced with a no-flux boundary generated by a conductive wall. The electric field can still penetrate this no-flux region due to the wall's finite conductivity, but ion migration is impeded due to the physicality of the wall. We perform detailed concentration and separation experiments across varying electric potentials, flow rates, and TE concentrations. We also show that TPE can achieve a 60,000-fold concentration factor continuously without an LE, using only 10 V DC. In comparison with conventional batch-driven ITP, continuous free-flow wall TPE (FFTPE) has the potential to serve as a simplified alternative method. FFTPE offers a high concentration power at a fraction of the required voltage, does not require an LE, and has the increased throughput potential of a continuous process.

Piezoresistive Conductive Microfluidic Membranes for Low-Cost On-Chip Pressure and Flow Sensing.

Over the last two decades, the field of microfluidics has received significant attention from both academia and industry. Each year, researchers report thousands of new prototype devices for use in a broad range of environmental, pharmaceutical, and biomedical engineering applications. While lab-on-a-chip fabrication costs have continued to decrease, the hardware required for monitoring fluid flows within the microfluidic devices themselves remains expensive and often cost-prohibitive for researchers interested in starting a microfluidics project. As microfluidic devices become capable of handling complex fluidic systems, low-cost, precise, and real-time pressure and flow rate measurement capabilities have become increasingly important. While many labs use commercial platforms and sensors, these solutions can often cost thousands of dollars and can be too bulky for on-chip use. Here we present a new inexpensive and easy-to-use piezoresistive pressure and flow sensor that can be easily integrated into existing on-chip microfluidic channels. The sensor consists of PDMS-carbon black conductive membranes and uses an impedance analyzer to measure impedance changes due to fluid pressure. The sensor costs several orders of magnitude less than existing commercial platforms and can monitor local fluid pressures and calculate flow rates based on the pressure gradient.

Correction: Microfluidic pumping, routing and metering by contactless metal-based electro-osmosis.

Correction for 'Microfluidic pumping, routing and metering by contactless metal-based electro-osmosis' by Xiaotong Fu et al., Lab Chip, 2015, DOI: 10.1039/c5lc00504c.

Microfluidic pumping, routing and metering by contactless metal-based electro-osmosis.

Over the past decade, many microfluidic platforms for fluid processing have been developed in order to perform on-chip fluidic manipulations. Many of these methods, however, require expensive and bulky external supporting equipment, which are not typically applicable for microsystems requiring portability. We have developed a new type of portable contactless metal electro-osmotic micropump capable of on-chip fluid pumping, routing and metering. The pump operates using two pairs of gallium metal electrodes, which are activated using an external voltage source, and separated from a main flow channel by a thin micron-scale PDMS membrane. The thin contactless membrane allows for field penetration and electro-osmotic (EO) flow within the microchannel, but eliminates electrode damage and sample contamination commonly associated with traditional DC electro-osmotic pumps that utilize electrodes in direct contact with the working fluid. The maximum flow rates and pressures generated by the pump using DI water as a working buffer are 10 nL min(-1) and 30 Pa, respectively. With our current design, the maximum operational conductivity where fluid flow is observed is 0.1 mS cm(-1). Due to the small size and simple fabrication procedure, multiple micropump units can be integrated into a single microfluidic device for automated on-chip routing and sample metering applications. We experimentally demonstrated the ability to quantify micropump electro-osmotic flowrate and pressure as a function of applied voltage, and developed a mathematical model capable of predicting the performance of a contactless micropump for a given external load and internal hydrodynamic microchannel resistance. Finally, we showed that by activating specific pumps within a microchannel network, our micropumps are capable of routing microchannel fluid flow and generating plugs of solute.