Analysis of pressure-driven membrane preconcentration for point-of-care assays.

Point-of-care diagnostic devices for both physicians and patients themselves are now ubiquitous, but often not sensitive enough for highly dilute analytes (e.g., pre-symptomatic viral detection). Two primary methods to address this challenge include (1) increasing the sensitivity of molecular recognition elements with greater binding affinity to the analyte or (2) increasing the concentration of the analyte being detected in the sample itself (preconcentration). The latter approach, preconcentration, is arguably more attractive if it can be made universally applicable to a wide range of analytes. In this study, pressure-driven membrane preconcentration devices were developed, and their performance was analyzed for detecting target analytes in biofluids in the form of point-of-care lateral-flow assays (LFAs). The demonstrated prototypes utilize negative or positive pressure gradients to move both water and small interferents (salt, pH) through a membrane filter, thereby concentrating the analyte of interest in the remaining sample fluid. Preconcentration up to 33× is demonstrated for influenza A nucleoprotein with a 5 kDa pore polyethersulfone membrane filter. LFA results are obtained within as short as several minutes and device operation is simple (very few user steps), suggesting that membrane preconcentration can be preferable to more complex and slow conventional preconcentration techniques used in laboratory practice.

Ultra-simple wearable local sweat volume monitoring patch based on swellable hydrogels.

Quantifiably monitoring sweat rate and volume is important to assess the stress level of individuals and/or prevent dehydration, but despite intense research, a convenient, continuous, and low-cost method to monitor sweat rate and total sweat volume loss remains an un-met need. We present here an ultra-simple wearable sensor capable of measuring sweat rate and volume accurately. The device continuously monitors sweat rate by wicking the produced sweat into hydrogels that measurably swell in their physical geometry. The device has been designed as a simple to fabricate, low-cost, disposable patch. This patch exhibits stable and predictable operation over the maximum variable chemistry expected for sweat (pH 4-9 and salinity 0-100 mM NaCl). Preliminary in vivo testing of the patch has been achieved during aerobic exercise, and the sweat rates measured via the patch accurately follow actual sweat rates.

Complete validation of a continuous and blood-correlated sweat biosensing device with integrated sweat stimulation.

A wearable sweat biosensing device is demonstrated that stimulates sweat and continuously measures sweat ethanol concentrations at 25 s intervals, which is then correlated with blood ethanol during a >3 hour testing phase. The testing involves a baseline condition (no ethanol) followed by a rapid blood and sweat rise of ethanol (oral bolus), and finally, the physiological response of the body as ethanol concentrations return to baseline (metabolized). Data sets include multiple in vivo validation trials and careful in vitro characterization of the electrochemical enzymatic ethanol sensor against likely interferents. Furthermore, the data is analyzed through known pharmacokinetic models with a strong linear Pearson correlation of 0.9474-0.9996. The continuous nature of the data also allows analysis of blood-to-sweat lag times that range between 2.3 to 11.41 min for ethanol signal onset and 19.32 to 34.44 min for the overall pharmacokinetic curve lag time. This work represents a significant advance that builds upon a continuum of previous work. However, unresolved questions include operation for 24 hours or greater and with analytes beyond those commonly explored for sweat (electrolytes and metabolites). Regardless, this work validates that sweat biosensing can provide continuous and blood-correlated data in an integrated wearable device.

Open nanofluidic films with rapid transport and no analyte exchange for ultra-low sample volumes.

Moving to ultra-low (<100 nL) sample volumes presents numerous challenges, many of which can be resolved by implementation of open nanofluidic films. These nanofluidic films are fabricated using a hexagonal network of gold-coated open microchannels which capture all of the following innovative advantages: (1) sample volumes of <100 nL cm-2; (2) zero analyte exchange and loss with the film materials; (3) rapid and omni-directional wicking transport of >500 nL min-1 per square of film; (4) ultra-simple roll-to-roll fabrication; (5) stable and bio-compatible super-hydrophilicity for weeks in air by peptide surface modification. Validation includes both detailed in vitro characterization and in vivo validation with sweat transport from the human skin. Sampling times (skin-to-sensor) of <3 min were achieved, setting new benchmarks for the field of wearable sweat sensing. This work addresses significant challenges for sweat biosensing, or for any other nano-liter regime (<100 nL) fluid sampling and sensing application.

Membrane isolation of repeated-use sweat stimulants for mitigating both direct dermal contact and sweat dilution.

With the device integration of sweat stimulation, sweat becomes a stronger candidate for non-invasive continuous biochemical sensing. However, sweat stimulants are cholinergenic agents and non-selective to just the sweat glands, and so, direct placement of sweat stimulants poses additional challenges in the possibility for uncontrollable transport of the stimulant into the body and challenges in contamination of the sweat sample. Reported here is membrane isolation of repeated-use sweat stimulants for mitigating direct dermal contact, dilution of the sweat stimulant, and contamination of the sweat sample. The membrane dramatically reduces passive diffusion of the sweat stimulant carbachol by roughly two orders of magnitude, while still allowing repeated sweat stimulation by iontophoretic delivery of the carbachol through the membrane and into the skin. Both in-vivo and in-vitro validation reveal feasibility for reliable integration of sweat stimulants within a wearable device for use periods of 24 h or more. In addition, advanced topics and confounding issues such as stimulant gel design, osmotic pressure, and ionic impurities are speculatively and theoretically discussed.

Wearable sensors: modalities, challenges, and prospects.

Wearable sensors have recently seen a large increase in both research and commercialization. However, success in wearable sensors has been a mix of both progress and setbacks. Most of commercial progress has been in smart adaptation of existing mechanical, electrical and optical methods of measuring the body. This adaptation has involved innovations in how to miniaturize sensing technologies, how to make them conformal and flexible, and in the development of companion software that increases the value of the measured data. However, chemical sensing modalities have experienced greater challenges in commercial adoption, especially for non-invasive chemical sensors. There have also been significant challenges in making significant fundamental improvements to existing mechanical, electrical, and optical sensing modalities, especially in improving their specificity of detection. Many of these challenges can be understood by appreciating the body's surface (skin) as more of an information barrier than as an information source. With a deeper understanding of the fundamental challenges faced for wearable sensors and of the state-of-the-art for wearable sensor technology, the roadmap becomes clearer for creating the next generation of innovations and breakthroughs.

Superwetting and aptamer functionalized shrink-induced high surface area electrochemical sensors.

Electrochemical sensing is moving to the forefront of point-of-care and wearable molecular sensing technologies due to the ability to miniaturize the required equipment, a critical advantage over optical methods in this field. Electrochemical sensors that employ roughness to increase their microscopic surface area offer a strategy to combatting the loss in signal associated with the loss of macroscopic surface area upon miniaturization. A simple, low-cost method of creating such roughness has emerged with the development of shrink-induced high surface area electrodes. Building on this approach, we demonstrate here a greater than 12-fold enhancement in electrochemically active surface area over conventional electrodes of equivalent on-chip footprint areas. This two-fold improvement on previous performance is obtained via the creation of a superwetting surface condition facilitated by a dissolvable polymer coating. As a test bed to illustrate the utility of this approach, we further show that electrochemical aptamer-based sensors exhibit exceptional signal strength (signal-to-noise) and excellent signal gain (relative change in signal upon target binding) when deployed on these shrink electrodes. Indeed, the observed 330% gain we observe for a kanamycin sensor is 2-fold greater than that seen on planar gold electrodes.

A new oil/membrane approach for integrated sweat sampling and sensing: sample volumes reduced from µL's to nL's and reduction of analyte contamination from skin.

Wearable sweat biosensensing technology has dominantly relied on techniques which place planar-sensors or fluid-capture materials directly onto the skin surface. This 'on-skin' approach can result in sample volumes in the µL regime, due to the roughness of skin and/or due to the presence of hair. Not only does this increase the required sampling time to 10's of minutes or more, but it also increases the time that sweat spends on skin and therefore increases the amount of analyte contamination coming from the skin surface. Reported here is a first demonstration of a new paradigm in sweat sampling and sensing, where sample volumes are reduced from the µL's to nL's regime, and where analyte contamination from skin is reduced or even eliminated. A micro-porous membrane is constructed such that it is porous to sweat only. To complete a working device, first placed onto skin is a cosmetic-grade oil, secondly this membrane, and thirdly the sensors. As a result, spreading of sweat is isolated to only regions above the sweat glands before it reaches the sensors. Best case sampling intervals are on the order of several minutes, and the majority of hydrophilic (low oil solubility) contaminants from the skin surface are blocked. In vitro validation of this new approach is performed with an improved artificial skin including human hair. In vivo tests show strikingly consistent results, and reveal that the oil/membrane is robust enough to even allow horizontal sliding of a sensor.

Electrokinetic pixels with biprimary inks for color displays and color-temperature-tunable smart windows.

We report on the advanced implementation of the biprimary color system in applications where subtractive color is performed inside a single pixel to alter the magnitude and color of reflection (electronic paper displays) or the optical transmission and color temperature (smart windows). A novel device structure can switch between four states: clear, black, either of two complementary colors from RGB and CMY sets, and also mixed states between one of these four states. The device structure utilizes an electrokinetic pixel structure, which combines the spectral performance of in-plane electrophoretic devices with the improved switching speeds of vertical electrophoresis. The electrophoretic dispersions are dual-particle dual-colored and are controlled using two traditional planar electrokinetic electrodes on the front and back substrates, along with a third electrode conveniently located at the perimeter of each unit cell. Demonstrated performance includes contrast ratios reaching ~10∶1, reflectance of ~62%, and transparency of ~75%. For electronic paper displays, these results provide a pathway to double the reflective performance compared to the traditional RGBW color-filter approach. For smart windows, the technology provides not only control of shade (transmission) but also provides complete control over color temperature. Furthermore, this three-electrode device can be roll-to-roll fabricated without need for any alignment steps, requiring only a single micro-replication step followed by self-aligned contact printing of the third electrode.

The microfluidics of the eccrine sweat gland, including biomarker partitioning, transport, and biosensing implications.

Non-invasive and accurate access of biomarkers remains a holy grail of the biomedical community. Human eccrine sweat is a surprisingly biomarker-rich fluid which is gaining increasing attention. This is especially true in applications of continuous bio-monitoring where other biofluids prove more challenging, if not impossible. However, much confusion on the topic exists as the microfluidics of the eccrine sweat gland has never been comprehensively presented and models of biomarker partitioning into sweat are either underdeveloped and/or highly scattered across literature. Reported here are microfluidic models for eccrine sweat generation and flow which are coupled with review of blood-to-sweat biomarker partition pathways, therefore providing insights such as how biomarker concentration changes with sweat flow rate. Additionally, it is shown that both flow rate and biomarker diffusion determine the effective sampling rate of biomarkers at the skin surface (chronological resolution). The discussion covers a broad class of biomarkers including ions (Na(+), Cl(-), K(+), NH4 (+)), small molecules (ethanol, cortisol, urea, and lactate), and even peptides or small proteins (neuropeptides and cytokines). The models are not meant to be exhaustive for all biomarkers, yet collectively serve as a foundational guide for further development of sweat-based diagnostics and for those beginning exploration of new biomarker opportunities in sweat.

Experimental and numerical insights into isotropic spreading and deterministic dewetting of dielectrowetted films.

Dielectrowetting effects of surface wrinkling, isotropic vs anisotropic spreading, electrode geometry, and deterministic dewetting are presented both experimentally and by 3D numerical modeling. The numerical results are generated by COMSOL in conjunction with the phase-field and electrohydrodynamic methods, including comparisons to experimental data. The dynamic behavior of the two-phase system has been accurately characterized on both the macro- and microscopic level. This work provides a deeper theoretical insight into the operating physics of dielectrowetting superspreading devices.

Investigation of Laplace barriers for arrayed electrowetting lab-on-a-chip.

Partial-post Laplace barriers have been postulated as a means to allow electrowetting transport and geometrical reshaping of fluids, followed by the preservation of fluid geometry after the electrowetting voltage is removed. Reported here is the first investigation of Laplace barriers with the arrayed electrodes and splitting/merging transport functions for an electrowetting lab-on-a-chip. Laplace barriers optimized for 500 × 500 µm(2) electrodes and 78 µm channel height are shown to provide geometrical control of fluid shape down to radii of curvature of ~70 µm. The Laplace barriers increase the splitting volume error, but with proper electrical control, the average error in the split volume is reduced to 5%. Improved programmable fluid storage in droplets or reservoirs and continuous channel flow are also shown. This work confirms the potential benefits of Laplace barriers for lab-on-a-chip and also reveals the unique challenges and operation requirements for Laplace barriers in lab-on-a-chip applications.

Scaling dielectrowetting optical shutters to higher resolution: microfluidic and optical implications.

A detailed study is reported on the implications of scaling dielectrowetting optical shutters to higher resolutions. Reducing droplet sizes from millimeters to 100 µm in diameter increases the relevance of microfluidic physics such as pinning, film breakup, and dewetting speed as well as optical physics such as transmission and diffraction. In addition, in this work we present improved material systems, including optimized dielectric stacks which reduce electrochemical degradation, and blended lower-viscosity fluids which increase dewetting speed. A higher-resolution device of ~250 µm diameter demonstrates switching speeds of <100 ms and a clear, optically transmissive aperture of >70%. In addition to revealing science not previously discussed, this work has strong applied importance as scaling to higher resolutions is desirable for improving visual appearance in applications ranging from smart windows to electronic signage.

Bright e-Paper by transport of ink through a white electrofluidic imaging film.

Many of the highest performance approaches for electronic paper use voltage to reveal or hide dark pigments or dyes over a white pixel surface, and the reflectance of white pixels is lower than in real paper because the dark pigments or dyes can never be fully removed from the visible pixel area. Here, we introduce a re-designed approach for electronic paper that transposes coloured ink in front of or behind a white microfluidic film. Pixels can provide >90% reflective area and have demonstrated <15 ms switching for 150 pixels-per-inch resolution. This new approach is also the first of its kind for electrowetting-style displays by allowing non-aligned lamination fabrication, and is the first ever colourant-transposing pixel that eliminates the need for ink microencapsulation or pixel borders.

Investigation of five types of switchable retroreflector films for enhanced visible and infrared conspicuity applications.

We report on the physics, design, characterization, and demonstration of five viable techniques for switchable retroreflectors, including integrated electrowetting scattering, integrated and external electrowetting light valves, external liquid crystal light valve, and external liquid crystal scattering. All techniques were evaluated for use in conspicuity applications spanning wavelengths in the visible and IR (night vision). Achieved performance includes high optical efficiencies up to nearly 30% (out of a maximum 35%), visibly fast switching speeds of <100 ms, low to moderate operating voltages ranging from 5 to 60 V, more than ±45 deg of operation angle, and implementation with pressure-sensitive, adhesive-backed films of 0.7 to 1 mm thickness for flexibility and impact resistance. Each approach has unique strengths and weaknesses, which will also be discussed for applications ranging from commercial to military conspicuity.

Partial-post Laplace barriers for virtual confinement, stable displacement, and >5 cm s(-1) electrowetting transport.

Laplace barriers composed of full-posts or ridges have been previously reported as a mechanism for virtual fluid confinement, but with unstable displacement (capillary fingering or fluid trapping, respectively). A new platform of 'partial-posts' eliminates the disadvantages of full-posts or ridges, while providing ~60-80% open channel area for rapid electrowetting fluid transport (>5 cm s(-1)). The fluid mechanics of partial-post Laplace barriers are far more complex than previous Laplace barriers as it involves two mechanisms: fluid can first begin to propagate either between, or under, the partial-posts. Careful design of channel and partial-post geometries is required, else one mechanism will dominate over the other. The physics and performance of partial-post Laplace barriers are verified using theoretical equations, experimental results, and dynamic numerical modeling.

Laplace barriers for electrowetting thresholding and virtual fluid confinement.

Reported are Laplace barriers consisting of arrayed posts or ridges that impart ∼100 to 1000 s of N/m(2) Laplace pressure for fluid confinement, but the Laplace pressure is also small enough such that the barriers are porous to electrowetting control. As a result, the barriers are able to provide electrowetting flow thresholding and virtual fluid confinement in noncircular fluid geometries. A simple theoretical model for the barriers and experimental demonstrations validate functionality that may be useful for lab-on-chip, display devices, and passive matrix control, to name a few applications.

Preparation and analysis of 1-chloronaphthalene for highly refractive electrowetting optics.

High-index oils are critical to the performance of optical electrowetting devices, such as lenses, prisms, and retroreflectors. Herein, the preparation and electrowetting analysis of 1-chloronaphthalene are reported. When 1-chloronaphthalene is mixed with small amounts of an alkane, the following properties can be achieved: refractive index > 1.60, viscosity < 5 cP for rapid switching, operation at <10 V, and +/-45 degrees of electrowetting modulation around the condition for a flat meniscus (90 degrees ).