Paul Yager

Microfluidic diagnostic technologies for global public health

The developing world does not have access to many of the best medical diagnostic technologies; they were designed for air-conditioned laboratories, refrigerated storage of chemicals, a constant supply of calibrators and reagents, stable electrical power, highly trained personnel and rapid transportation of samples. Microfluidic systems allow miniaturization and integration of complex functions, which could move sophisticated diagnostic tools out of the developed-world laboratory. These systems must be inexpensive, but also accurate, reliable, rugged and well suited to the medical and social contexts of the developing world.

Point-of-Care Diagnostics for Global Health

Biomedical engineers have traditionally developed technologies in response to the needs of the developed worlds medical community. As a result, the diagnostic systems on which they have worked have met the requirements of well-funded laboratories in highly regulated and quality-assessed environments. However, such approaches do not address the needs of the majority of the worlds people afflicted with infectious diseases, who have, at best, access to poorly resourced health care facilities with almost no supporting clinical laboratory infrastructure. A major challenge for the biomedical engineering community is to develop diagnostic tests to meet the needs of these people, the majority of whom are in the developing world. We here review the context in which the diagnostics must operate, some of the appropriate diagnostic technologies already in distribution, and some emerging technologies that promise to address this challenge. However, there is much room for innovation, adaptation, and cost reduction before these technologies can impact health care in the developing world.

Biotechnology at low Reynolds numbers.

The shrinking of liquid handling systems to the micron and submicron size range entails moving into the area of small Reynolds numbers. The fluid dynamics in this regime are very different from the macroscale. We present an intuitive explanation of how the different physics of small Reynolds numbers flow, along with microscopic sizes, can influence device design, and give examples from our own work using fluid flow in microfabricated devices designed for biological processing.

A rapid diffusion immunoassay in a T-sensor

We have developed a rapid diffusion immunoassay that allows measurement of small molecules down to subnanomolar concentrations in <1 min. This competitive assay is based on measuring the distribution of a labeled probe molecule after it diffuses for a short time from one region into another region containing antigen-specific antibodies. The assay was demonstrated in the T-sensor, a simple microfluidic device that places two fluid streams in contact and allows interdiffusion of their components. The model analyte was phenytoin, a typical small drug molecule. Clinically relevant levels were measured in blood diluted from 10- to 400-fold in buffer containing the labeled antigen. Removal of cells from blood samples was not necessary. This assay compared favorably with fluorescence polarization immunoassay (FPIA) measurements. Numerical simulations agree well with experimental results and provide insight for predicting assay performance and limitations. The assay is homogeneous, requires <1 μl of reagents and sample, and is applicable to a wide range of analytes.

A ferrofluidic magnetic micropump

A microfluidic pump is described that uses magnetic actuation to push fluid through a microchannel. Operation relies on the use of magnetically-actuated plugs of ferrofluid, a suspension of nanosize ferromagnetic particles. The ferrofluid contacts but is immiscible with the pumped fluid. The prototype circular design demonstrates continuous pumping by regenerating a translating ferrofluidic plug at the conclusion of each pumping cycle. The flow rate can be controlled by adjusting device dimensions or the velocity of an external permanent magnet that directs the motion of the ferrofluid. The ferrofluidic plugs also serve as valves; if the magnetic actuator is stopped, pressure can be maintained with no power consumption. Flow can also be reversed by switching the direction of actuation. The maximum flow rate achieved with minimal backpressure was 45.8 /spl mu/l/min. The maximum pressure head achieved was 135 mm water (1.2 kPa).

Controlled reagent transport in disposable 2D paper networks.

Recent reports have demonstrated the multi-analyte detection capability of paper networks with multiple outlets per inlet. In this report, we focus on the capabilities of 2D paper networks with multiple inlets per outlet and demonstrate the controlled transport of reagents within paper devices. Specifically, we demonstrate methods of controlling fluid transport using the geometry of the network and dissolvable barriers. Finally, we discuss the implications for higher sensitivity detection using this type of 2D paper network.

Microfluidics without pumps: reinventing the T-sensor and H-filter in paper networks.

Conventional microfluidic devices typically require highly precise pumps or pneumatic control systems, which add considerable cost and the requirement for power. These restrictions have limited the adoption of microfluidic technologies for point-of-care applications. Paper networks provide an extremely low-cost and pumpless alternative to conventional microfluidic devices by generating fluid transport through capillarity. We revisit well-known microfluidic devices for hydrodynamic focusing, sized-based extraction of molecules from complex mixtures, micromixing, and dilution, and demonstrate that paper-based devices can replace their expensive conventional microfluidic counterparts.

Diffusion-based extraction in a microfabricated device

Abstract Microfabricated fluid systems allow complex chemical analyses to be performed on sub-nanoliter volumes. However, many common laboratory procedures, including filtration, have yet to be robustly implemented in micro-fluid systems. A device has been developed to separate particles and molecules based on their diffusion coefficients; the process is demonstrated using a micromachined device with fluid channels as small as 20 μm. A simple model predicts exponential dependence of the output concentration on diffusion coefficient in certain regimes. Experiments confirm the model.

Theoretical Analysis of Molecular Diffusion in Pressure-Driven Laminar Flow in Microfluidic Channels

The T-sensor is a microfluidic analytical device that operates at low Reynolds numbers to ensure entirely laminar flow. Diffusion of molecules between streams flowing side by side may be observed directly. The pressure-driven velocity profile in the duct-shaped device influences diffusive transport in ways that affect the use of the T-sensor to measure molecular properties. The primary effect is a position-dependent variation in the extent of diffusion that occurs due to the distribution of residence time among different fluid laminae. A more detailed characterization reveals that resultant secondary concentration gradients yield variations in the scaling behavior between diffusive displacement and elapsed time in different regions of the channel. In this study, the time-dependent evolution of analyte distribution has been quantified using a combination of one- and two-dimensional models. The results include an accurate portrayal of the shape of the interdiffusion region in a representative T-sensor assay, calculation of the diffusive scaling law across the width of the channel, and quantification of artifacts that occur when making diffusion coefficient measurements in the T-sensor.

Optical Measurement of Transverse Molecular Diffusion in a Microchannel

Quantitative analysis of molecular diffusion is a necessity for the efficient design of most microfluidic devices as well as an important biophysical method in its own right. This study demonstrates the rapid measurement of diffusion coefficients of large and small molecules in a microfluidic device, the T-sensor, by means of conventional epifluorescence microscopy. Data were collected by monitoring the transverse flux of analyte from a sample stream into a second stream flowing alongside it. As indicated by the low Reynolds numbers of the system (< 1), flow is laminar, and molecular transport between streams occurs only by diffusion. Quantitative determinations were made by fitting data with predictions of a one-dimensional model. Analysis was made of the flow development and its effect on the distribution of diffusing analyte using a three-dimensional modeling software package. Diffusion coefficients were measured for four fluorescently labeled molecules: fluorescein-biotin, insulin, ovalbumin, and streptavidin. The resulting values differed from accepted results by an average of 2.4%. Microfluidic system parameters can be selected to achieve accurate diffusion coefficient measurements and to optimize other microfluidic devices that rely on precise transverse transport of molecules.