Sub-micrometer-precision, three-dimensional (3D) hydrodynamic focusing via "microfluidic drifting".

In this article, we demonstrate single-layered, "microfluidic drifting" based three-dimensional (3D) hydrodynamic focusing devices with particle/cell focal positioning approaching submicron precision along both lateral and vertical directions. By systematically optimizing channel geometries and sample/sheath flow rates, a series of "microfluidic drifting" based 3D hydrodynamic focusing devices with different curvature angles are designed and fabricated. Their performances are then evaluated using confocal microscopy, fast camera imaging, and side-view imaging techniques. Using a device with a curvature angle of 180°, we have achieved a standard deviation of ±0.45 µm in particle focal position and a coefficient of variation (CV) of 2.37% in flow cytometric measurements. To the best of our knowledge, this is the best CV that has been achieved using a microfluidic flow cytometry device. Moreover, the device showed the capability to distinguish 8 peaks when subjected to a stringent 8-peak rainbow calibration test, signifying the ability to perform sensitive, accurate tests similar to commercial flow cytometers. We have further tested and validated our device by detection of HEK-293 cells. With its advantages in simple fabrication (i.e., single-layered device), precise 3D hydrodynamic focusing (i.e., submicrometer precision along both lateral and vertical directions), and high detection resolution (i.e., low CV), our method could serve as an important basis for high-performance, mass-producible microfluidic flow cytometry.

Surface acoustic wave microfluidics.

The recent introduction of surface acoustic wave (SAW) technology onto lab-on-a-chip platforms has opened a new frontier in microfluidics. The advantages provided by such SAW microfluidics are numerous: simple fabrication, high biocompatibility, fast fluid actuation, versatility, compact and inexpensive devices and accessories, contact-free particle manipulation, and compatibility with other microfluidic components. We believe that these advantages enable SAW microfluidics to play a significant role in a variety of applications in biology, chemistry, engineering and medicine. In this review article, we discuss the theory underpinning SAWs and their interactions with particles and the contacting fluids in which they are suspended. We then review the SAW-enabled microfluidic devices demonstrated to date, starting with devices that accomplish fluid mixing and transport through the use of travelling SAW; we follow that by reviewing the more recent innovations achieved with standing SAW that enable such actions as particle/cell focusing, sorting and patterning. Finally, we look forward and appraise where the discipline of SAW microfluidics could go next.

Unconventional microfluidics: expanding the discipline.

Since its inception, the discipline of microfluidics has been harnessed for innovations in the biomedicine/chemistry fields-and to great effect. This success has had the natural side-effect of stereotyping microfluidics as a platform for medical diagnostics and miniaturized lab processes. But microfluidics has more to offer. And very recently, some researchers have successfully applied microfluidics to fields outside its traditional domains. In this Focus article, we highlight notable examples of such "unconventional" microfluidics applications (e.g., robotics, electronics). It is our hope that these early successes in unconventional microfluidics prompt further creativity, and inspire readers to expand the microfluidics discipline.

Exploiting mechanical biomarkers in microfluidics.

Cellular mechanical properties have been observed to have important implications for pathogenesis and pathophysiology. These observations have led to the recent development of a unique class of biomarkers: mechanical biomarkers. Compared with the traditional biochemical-based biomarkers (e.g., antibodies), mechanical biomarkers have many advantages such as label-free, low cost, convenient maintenance, and reduced assay time. In the past few years, there has been an increasing effort to exploit cellular mechanical biomarkers in microfluidic devices. This trend makes sense because microfluidic devices often feature structures that have characteristic lengths similar to those of cells, which renders them uniquely capable of probing and utilizing mechanical biomarkers. In this Focus article, we discuss a few examples of mechanical biomarker-based microfluidic applications. We believe that these examples are just the tip of the iceberg and that the full potential of mechanical biomarkers in microfluidic-based diagnostics and therapeutics has yet to be revealed.

Surface acoustic wave (SAW) acoustophoresis: now and beyond.

On-chip manipulation of micro-objects has long been sought to facilitate fundamental biological studies and point-of-care diagnostic systems. In recent years, research on surface acoustic wave (SAW) based micro-object manipulation (i.e., SAW acoustophoresis) has gained significant momentum due to its many advantages, such as non-invasiveness, versatility, simple fabrication, easy operation, and convenient integration with other on-chip units. SAW acoustophoresis is especially useful for lab-on-a-chip applications where a compact and non-invasive biomanipulation technique is highly desired. In this Focus article, we discuss recent advancements in SAW acoustophoresis and provide some perspectives on the future development of this dynamic field.

Microfluidic synthesis of multifunctional Janus particles for biomedical applications.

Multifunctional Janus particles have a variety of applications in a wide range of fields. However, to achieve many of these applications, high-throughput, low-cost techniques are needed to synthesize these particles with precise control of the various structural/physical/chemical properties. Microfluidics provides a unique platform to fabricate Janus particles using carefully controlled liquid flow in microfluidic channels to form Janus droplets and various types of solidification methods to solidify them into Janus particles. In this Focus article, we summarize the most recent representative works on Janus particle fabrication in microfluidics. The applications of Janus particles in biomedical areas are emphasized. We believe that microfluidics-enabled multifunctional Janus particles could resolve multiple prevalent issues in biomedicine (e.g., disease monitoring at an early stage, high-throughput bioassays, therapeutic delivery) if persistent effort and collaboration are devoted to this direction.

An integrated, multiparametric flow cytometry chip using "microfluidic drifting" based three-dimensional hydrodynamic focusing.

In this work, we demonstrate an integrated, single-layer, miniature flow cytometry device that is capable of multi-parametric particle analysis. The device integrates both particle focusing and detection components on-chip, including a "microfluidic drifting" based three-dimensional (3D) hydrodynamic focusing component and a series of optical fibers integrated into the microfluidic architecture to facilitate on-chip detection. With this design, multiple optical signals (i.e., forward scatter, side scatter, and fluorescence) from individual particles can be simultaneously detected. Experimental results indicate that the performance of our flow cytometry chip is comparable to its bulky, expensive desktop counterpart. The integration of on-chip 3D particle focusing with on-chip multi-parametric optical detection in a single-layer, mass-producible microfluidic device presents a major step towards low-cost flow cytometry chips for point-of-care clinical diagnostics.

Microfluidic diagnostics for the developing world.

For more than a decade, it has been expected that microfluidic technology would revolutionize the healthcare industry with simple, inexpensive, effective, and ubiquitous miniature diagnostic devices. To date, however, microfluidics has not yet been able to live up to these expectations. This fact has led to the recent development of new philosophies and methodologies for microfluidic diagnostics. In this Focus article, we will discuss some of the latest breakthroughs that could significantly impact medical diagnostics in the developing world.

A single-layer, planar, optofluidic Mach-Zehnder interferometer for label-free detection.

We have developed a planar, optofluidic Mach-Zehnder interferometer for the label-free detection of liquid samples. In contrast to most on-chip interferometers which require complex fabrication, our design was realized via a simple, single-layer soft lithography fabrication process. In addition, a single-wavelength laser source and a silicon photodetector were the only optical equipment used for data collection. The device was calibrated using published data for the refractive index of calcium chloride (CaCl(2)) in solution, and the biosensing capabilities of the device were tested by detecting bovine serum albumin (BSA). Our design enables a refractometer with a low limit of detection (1.24 × 10(-4) refractive index units (RIU)), low variability (1 × 10(-4) RIU), and high sensitivity (927.88 oscillations per RIU). This performance is comparable to state-of-the-art optofluidic refractometers that involve complex fabrication processes and/or expensive, bulky optics. The advantages of our device (i.e. simple fabrication process, straightforward optical equipment, low cost, and high detection sensitivity) make it a promising candidate for future mass-producible, inexpensive, highly sensitive, label-free optical detection systems.

Tunable two-dimensional liquid gradient refractive index (L-GRIN) lens for variable light focusing.

We report a two-dimensional (2D) tunable liquid gradient refractive index (L-GRIN) lens for variable focusing of light in the out-of-plane direction. This lens focuses a light beam through a liquid medium with a 2D hyperbolic secant (HS) refractive index gradient. The refractive index gradient is established in a microfluidic chamber through the diffusion between two fluids with different refractive indices, i.e. CaCl(2) solution and deionized (DI) water. The 2D HS refractive index profile and subsequently the focal length of the L-GRIN lens can be tuned by changing the ratio of the flow rates of the CaCl(2) solution and DI water. The focusing effect is experimentally characterized through side-view and top-view image analysis, and the experimental data match well with the results from ray-tracing optical simulations. Advantages of the 2D L-GRIN lens include simple device fabrication procedure, low fluid consumption rate, convenient lens-tuning mechanism, and compatibility with existing microfluidic devices. We expect that with further optimizations, this 2D L-GRIN lens can be used in many optics-based lab-on-a-chip applications.

Optofluidic tunable microlens by manipulating the liquid meniscus using a flared microfluidic structure.

We have designed, demonstrated, and characterized a simple, novel in-plane tunable optofluidic microlens. The microlens is realized by utilizing the interface properties between two different fluids: CaCl(2)solution and air. A constant contact angle of ∼90° is the pivotal factor resulting in the outward bowing and convex shape of the CaCl(2) solution-air interface. The contact angle at the CaCl(2) solution-air interface is maintained by a flared structure in the polydimethylsiloxane channel. The resulting bowing interface, coupled with the refractive index difference between the two fluids, results in effective in-plane focusing. The versatility of such a design is confirmed by characterizing the intensity of a traced beam experimentally and comparing the observed focal points with those obtained via ray-tracing simulations. With the radius of curvature conveniently controlled via fluid injection, the resulting microlens has a readily tunable focal length. This ease of operation, outstandingly low fluid usage, large range tunable focal length, and in-plane focusing ability make this lens suitable for many potential lab-on-a-chip applications such as particle manipulation, flow cytometry, and in-plane optical trapping.

Acoustic tweezers: patterning cells and microparticles using standing surface acoustic waves (SSAW).

Here we present an active patterning technique named "acoustic tweezers" that utilizes standing surface acoustic wave (SSAW) to manipulate and pattern cells and microparticles. This technique is capable of patterning cells and microparticles regardless of shape, size, charge or polarity. Its power intensity, approximately 5x10(5) times lower than that of optical tweezers, compares favorably with those of other active patterning methods. Flow cytometry studies have revealed it to be non-invasive. The aforementioned advantages, along with this technique's simple design and ability to be miniaturized, render the "acoustic tweezers" technique a promising tool for various applications in biology, chemistry, engineering, and materials science.

A millisecond micromixer via single-bubble-based acoustic streaming.

We present ultra-fast homogeneous mixing inside a microfluidic channel via single-bubble-based acoustic streaming. The device operates by trapping an air bubble within a "horse-shoe" structure located between two laminar flows inside a microchannel. Acoustic waves excite the trapped air bubble at its resonance frequency, resulting in acoustic streaming, which disrupts the laminar flows and triggers the two fluids to mix. Due to this technique's simple design, excellent mixing performance, and fast mixing speed (a few milliseconds), our single-bubble-based acoustic micromixer may prove useful for many biochemical studies and applications.

Tunable Liquid Gradient Refractive Index (L-GRIN) lens with two degrees of freedom.

We report a tunable optofluidic microlens configuration named the Liquid Gradient Refractive Index (L-GRIN) lens for focusing light within a microfluidic device. The focusing of light was achieved through the gradient refractive index (GRIN) within the liquid medium, rather than via curved refractive lens surfaces. The diffusion of solute (CaCl(2)) between side-by-side co-injected microfluidic laminar flows was utilized to establish a hyperbolic secant (HS) refractive index profile to focus light. Tailoring the refractive index profile by adjusting the flow conditions enables not only tuning of the focal distance (translation mode), but also shifting of the output light direction (swing mode), a second degree of freedom that to our knowledge has yet to be accomplished for in-plane tunable microlenses. Advantages of the L-GRIN lens also include a low fluid consumption rate, competitive focusing performance, and high compatibility with existing microfluidic devices. This work provides a new strategy for developing integrative tunable microlenses for a variety of lab-on-a-chip applications.

Single-layer planar on-chip flow cytometer using microfluidic drifting based three-dimensional (3D) hydrodynamic focusing.

In this work, we demonstrate an on-chip microfluidic flow cytometry system based on a three-dimensional (3D) hydrodynamic focusing technique, microfluidic drifting. By inducing Dean flow in a curved microfluidic channel, microfluidic drifting can be used to hydrodynamically focus cells or particles in the vertical direction and enables the 3D hydrodynamic focusing in a single-layer planar microfluidic device. Through theoretical calculation, numerical simulation, and experimental characterization, we found that the microfluidic drifting technique can be effectively applied to three-dimensionally focus microparticles with density and size equivalent to those of human CD4+ T lymphocytes. In addition, we developed a flow cytometry platform by integrating the 3D focusing device with a laser-induced fluorescence (LIF) detection system. The system was shown to provide effective high-throughput flow cytometry measurements at a rate of greater than 1700 cells s(-1).

Focusing microparticles in a microfluidic channel with standing surface acoustic waves (SSAW).

We introduce a novel on-chip microparticle focusing technique using standing surface acoustic waves (SSAW). Our method is simple, fast, dilution-free, and applicable to virtually any type of microparticle.

"Microfluidic drifting"--implementing three-dimensional hydrodynamic focusing with a single-layer planar microfluidic device.

We introduce a novel fluid manipulation technique named "microfluidic drifting" to enable three-dimensional (3D) hydrodynamic focusing with a simple single-layer planar microfluidic device.

Hydrodynamically tunable optofluidic cylindrical microlens.

In this work, we report the design, fabrication, and characterization of a tunable optofluidic microlens that focuses light within a microfluidic device. The microlens is generated by the interface of two co-injected miscible fluids of different refractive indices, a 5 M CaCl(2) solution (n(D) = 1.445) and deionized (DI) water (n(D) = 1.335). When the liquids flow through a 90-degree curve in a microchannel, a centrifugal effect causes the fluidic interface to be distorted and the CaCl(2) solution bows outwards into the DI water portion. The bowed fluidic interface, coupled with the refractive index contrast between the two fluids, yields a reliable cylindrical microlens. The optical characteristics of the microlens are governed by the shape of the fluidic interface, which can be altered by simply changing the flow rate. Higher flow rates generate a microlens with larger curvature and hence shorter focal length. The changing of microlens profile is studied using both computational fluid dynamics (CFD) and confocal microscopy. The focusing effect is experimentally characterized through intensity measurements and image analysis of the focused light beam, and the experimental data are further confirmed by the results from a ray-tracing optical simulation. Our investigation reveals a simple, robust, and effective mechanism for integrating optofluidic tunable microlenses in lab-on-a-chip systems.

A nanoparticle amplification based quartz crystal microbalance DNA sensor for detection of Escherichia coli O157:H7.

A quartz crystal microbalance (QCM) DNA sensor, based on the nanoparticle amplification method, was developed for detection of Escherichia coli O157:H7. A thiolated single-stranded DNA (ssDNA) probe specific to E. coli O157:H7 eaeA gene was immobilized onto the QCM sensor surface through self-assembly. The hybridization was induced by exposing the ssDNA probe to the complementary target DNA, and resulted in the mass change and therefore frequency change of the QCM. Streptavidin conjugated Fe(3)O(4) nanoparticles (average diameter=145 nm) were used as "mass enhancers" to amplify the frequency change. Synthesized biotinylated oligonucleotides as well as E. coli O157:H7 eaeA gene fragments (151 bases) amplified using asymmetric PCR with biotin labeled primers were tested. As low as 10(-12)M synthesized oligonucleotides and 2.67 x 10(2) colony forming unit (CFU)/ml E. coli O157:H7 cells can be detected by the sensor. Linear correlation between frequency change and logarithmic number of bacterial cell concentration was found for E. coli O157:H7 from 2.67 x 10(2) to 2.67 x 10(6)CFU/ml.