Fluidic shaping and in-situ measurement of liquid lenses in microgravity.

In the absence of gravity, surface tension dominates over the behavior of liquids. While this often poses a challenge in adapting Earth-based technologies to space, it can also provide an opportunity for novel technologies that utilize its advantages. In particular, surface tension drives a liquid body to a constant-mean-curvature shape with extremely smooth surfaces, properties which are highly beneficial for optical components. We here present the design, implementation and analysis of parabolic flight experiments demonstrating the creation and in-situ measurement of optical lenses made entirely by shaping liquids in microgravity. We provide details of the two experimental systems designed to inject the precise amount of liquid within the short microgravity timeframe provided in a parabolic flight, while also measuring the resulting lens' characteristics in real-time using both resolution target-imaging and Shack-Hartmann wavefront sensing. We successfully created more than 20 liquid lenses during the flights. We also present video recordings of the process, from the lenses' creation during microgravity and up until their collapse upon return to gravity. The work thus demonstrates the feasibility of creating and utilizing liquid-based optics in space.

Dynamic control of high-voltage actuator arrays by light-pattern projection on photoconductive switches.

The ability to control high-voltage actuator arrays relies, to date, on expensive microelectronic processes or on individual wiring of each actuator to a single off-chip high-voltage switch. Here we present an alternative approach that uses on-chip photoconductive switches together with a light projection system to individually address high-voltage actuators. Each actuator is connected to one or more switches that are nominally OFF unless turned ON using direct light illumination. We selected hydrogenated amorphous silicon (a-Si:H) as our photoconductive material, and we provide a complete characterization of its light to dark conductance, breakdown field, and spectral response. The resulting switches are very robust, and we provide full details of their fabrication processes. We demonstrate that the switches can be integrated into different architectures to support both AC and DC-driven actuators and provide engineering guidelines for their functional design. To demonstrate the versatility of our approach, we demonstrate the use of the photoconductive switches in two distinctly different applications-control of µm-sized gate electrodes for patterning flow fields in a microfluidic chamber and control of cm-sized electrostatic actuators for creating mechanical deformations for haptic displays.

Automated device for multi-stage paper-based assays enabled by an electroosmotic pumping valve.

We leverage electroosmotic-flow generation in porous media in combination with a hydrophobic air gap to create a controllable valve capable of operating in either finite dosing or continuous flow mode, enabling the implementation of multi-step assays on paper-based devices. The hydrophobic air gap between two paper pads creates a barrier keeping the valve nominally closed. Electroosmotic actuation, implemented using a pair of electrodes under the upstream pad, generates sufficient pressure to overcome the barrier and connect the two pads. We present a model describing the flow and governing parameters, including the electric potentials required to open and close the valve and the threshold potential for switching between the modes of operation. We construct the air gap using a hierarchical superhydrophobic surface and study the stability of the closed valve under strenuous conditions and find good agreement between our model and experimental results, as well as stable working conditions for practical applications. We present a straightforward design for a compact and automated device based on paper pads placed on top of printed circuit boards (PCB), equipped with heating and actuation electrodes and additional power and logic capabilities. Finally, we demonstrate the use of the device for amplification of SARS-CoV-2 sequences directly from raw saliva samples, using a loop-mediated isothermal amplification (LAMP) protocol requiring sample lysis followed by enzymatic deactivation and delivery to multiple amplification sites. Since PCB costs scale favorably with mass-production, we believe that this approach could lead to a low-cost diagnostic device that offers the sensitivity of amplification methods.

Selective Extraction of Biomolecules Using a Bidirectional Flow Filter.

We present a microfluidic device for selective separation and extraction of molecules based on their diffusivity. The separation relies on electroosmotically driven bidirectional flows in which high-diffusivity species experience a net-zero velocity and lower diffusivity species are advected to a collection reservoir. The device can operate continuously and is suitable for processing low sample volumes. Using several model systems, we show that the extraction efficiency of the system is maintained at more than 90% over tens of minutes with a purity of more than 99%. We demonstrate the applicability of the device to the extraction of genomic DNA from short DNA fragments.

Reconfigurable microfluidics.

Lab-on-a-chip devices leverage microfluidic technologies to enable chemical and biological processes at small scales. However, existing microfluidic channel networks are typically designed for the implementation of a single function or a well-defined protocol and do not allow the flexibility and real-time experimental decision-making essential to many scientific applications. In this Perspective, we highlight that reconfigurability and programmability of microfluidic platforms can support new functionalities that are beyond the reach of current lab-on-a-chip systems. We describe the ideal fully reconfigurable microfluidic device that can change its shape and function dynamically, which would allow researchers to tune a microscale experiment with the capacity to make real-time decisions. We review existing technologies that can dynamically control microscale flows, suggest additional physical mechanisms that could be leveraged towards the goal of reconfigurable microfluidics and highlight the importance of these efforts for the broad scientific community.

Microscale Hydrodynamic Cloaking and Shielding via Electro-Osmosis.

We demonstrate theoretically and experimentally that injection of momentum in a region surrounding an object in microscale flow can yield both "cloaking" conditions, where the flow field outside the cloaking region is unaffected by the object, and "shielding" conditions, where the hydrodynamic forces on the object are eliminated. Using field-effect electro-osmosis as a mechanism for injection of momentum, we present a theoretical framework and analytical solutions for a range of geometrical shapes, validate these both numerically and experimentally, and demonstrate the ability to dynamically switch between the different states.

Microscopic scan-free surface profiling over extended axial ranges by point-spread-function engineering.

The shape of a surface, i.e., its topography, influences many functional properties of a material; hence, characterization is critical in a wide variety of applications. Two notable challenges are profiling temporally changing structures, which requires high-speed acquisition, and capturing geometries with large axial steps. Here, we leverage point-spread-function engineering for scan-free, dynamic, microsurface profiling. The presented method is robust to axial steps and acquires full fields of view at camera-limited framerates. We present two approaches for implementation: fluorescence-based and label-free surface profiling, demonstrating the applicability to a variety of sample geometries and surface types.

Microfluidic device for coupling isotachophoretic sample focusing with nanopore single-molecule sensing.

Solid-state nanopores (NPs) are label-free single-molecule sensors, capable of performing highly sensitive assays from a small number of biomolecule translocation events. However, single-molecule sensing is challenging at extremely low analyte concentrations due to the limited flux of analytes to the sensing volume. This leads to a low event rate and increases the overall assay time. In this work, we present a method to enhance the event rate at low analyte concentrations by using isotachophoresis (ITP) to focus and deliver analytes to a nanopore sensor. Central to this method is a device capable of performing ITP focusing directly on a solid-state NP chip, while preventing the focusing electric field from damaging the nanopore membrane. We discuss considerations and trade-offs related to the design of the focusing channel, the ITP electrolyte system and electrical decoupling between the focusing and sensing modes. Finally, we demonstrate an integrated device wherein the concentration enhancement due to ITP focusing leads to an increase in event rate of >300-fold in the ITP-NP device as compared to the NP-only case.

Intermediate States of Wetting on Hierarchical Superhydrophobic Surfaces.

Wetting transition on superhydrophobic surfaces is commonly described as an abrupt jump between two stable states-either from Cassie to Wenzel for nonhierarchical surfaces or from Cassie to nano-Cassie on hierarchical surfaces. We here experimentally study the electrowetting of hierarchical superhydrophobic surfaces composed of multiple length scales by imaging the light reflections from the gas-liquid interface. We present the existence of a continuous set of intermediate states of wetting through which the gas-liquid interface transitions under a continuously increasing external forcing. This transition is partially reversible and is limited only by localized Cassie to Wenzel transitions at nanodefects in the structure. In addition, we show that even a surface containing many localized wetted regions can still exhibit extremely low contact angle hysteresis, thus remaining useful for many heat transfer and self-cleaning applications. Expanding the classical definition of the Cassie state in the context of hierarchical surfaces, from a single state to a continuum of metastable states ranging from the centimeter to the nanometer scale, is important for a better description of the slip properties of superhydrophobic surfaces and provides new considerations for their effective design.

Tunable Bidirectional Electroosmotic Flow for Diffusion-Based Separations.

We present a new concept for on-chip separation that leverages bidirectional flow, to tune the dispersion regime of molecules and particles. The system can be configured so that low diffusivity species experience a ballistic transport regime and are advected through the chamber, whereas high diffusivity species experience a diffusion dominated regime with zero average velocity and are retained in the chamber. We detail the means of achieving bidirectional electroosmotic flow using an array of alternating current (AC) field-effect electrodes, experimentally demonstrate the separation of particles and antibodies from dyes, and present a theoretical analysis of the system, providing engineering guidelines for its design and operation.

Nonuniform Electro-osmotic Flow Drives Fluid-Structure Instability.

We demonstrate the existence of a fluid-structure instability arising from the interaction of electro-osmotic flow with an elastic substrate. Considering the case of flow within a soft fluidic chamber, we show that above a certain electric field threshold, negative gauge pressure induced by electro-osmotic flow causes the collapse of its elastic walls. We combine experiments and theoretical analysis to elucidate the underlying mechanism for instability and identify several distinct dynamic regimes. The understanding of this instability is important for the design of electrokinetic systems containing soft elements.

Electrokinetic Scanning Probe.

The theoretical analysis and experimental demonstration of a new concept are presented for a non-contact scanning probe, in which transport of fluid and molecules is controlled by electric fields. The electrokinetic scanning probe (ESP) enables local chemical and biochemical interactions with surfaces in liquid environments. The physical mechanism and design criteria for such a probe are presented, and its compatibility with a wide range of processing solutions and pH values are demonstrated. The applicability of the probe is shown for surface patterning in conjunction with localized heating and 250-fold analyte stacking.

Spatially Resolved Genetic Analysis of Tissue Sections Enabled by Microscale Flow Confinement Retrieval and Isotachophoretic Purification.

We have developed a method for spatially resolved genetic analysis of formalin-fixed paraffin-embedded (FFPE) cell block and tissue sections. This method involves local sampling using hydrodynamic flow confinement of a lysis buffer, followed by electrokinetic purification of nucleic acids from the sampled lysate. We characterized the method by locally sampling an array of points with a circa 200 µm diameter footprint, enabling the detection of single KRAS and BRAF point mutations in small populations of RKO and MCF-7 FFPE cell blocks. To illustrate the utility of this approach for genetic analysis, we demonstrate spatially resolved genotyping of FFPE sections of human breast invasive ductal carcinoma.

Electroosmotic Flow Dipole: Experimental Observation and Flow Field Patterning.

We experimentally demonstrate the phenomenon of electroosmotic dipole flow that occurs around a localized surface charge region under the application of an external electric field in a Hele-Shaw cell. We use localized deposition of polyelectrolytes to create well-controlled surface charge variations, and show that, for a disk-shaped spot, the internal pressure distribution that arises results in uniform flow within the spot and dipole flow around it. We further demonstrate the superposition of surface charge spots to create complex flow patterns, without the use of physical walls.

Dynamic microscale flow patterning using electrical modulation of zeta potential.

The ability to move fluids at the microscale is at the core of many scientific and technological advancements. Despite its importance, microscale flow control remains highly limited by the use of discrete channels and mechanical valves, and relies on fixed geometries. Here we present an alternative mechanism that leverages localized field-effect electroosmosis to create dynamic flow patterns, allowing fluid manipulation without the use of physical walls. We control a set of gate electrodes embedded in the floor of a fluidic chamber using an ac voltage in sync with an external electric field, creating nonuniform electroosmotic flow distributions. These give rise to a pressure field that drives the flow throughout the chamber. We demonstrate a range of unique flow patterns that can be achieved, including regions of recirculating flow surrounded by quiescent fluid and volumes of complete stagnation within a moving fluid. We also demonstrate the interaction of multiple gate electrodes with an externally generated flow field, allowing spatial modulation of streamlines in real time. Furthermore, we provide a characterization of the system in terms of time response and dielectric breakdown, as well as engineering guidelines for its robust design and operation. We believe that the ability to create tailored microscale flow using solid-state actuation will open the door to entirely new on-chip functionalities.

Dynamic control of capillary flow in porous media by electroosmotic pumping.

Microfluidic paper-based analytical devices (µPADs) rely on capillary flow to achieve filling, mixing and delivery of liquids. We investigate the use of electroosmotic (EO) pumping as a mechanism for dynamic control of capillary flow in paper-based devices. The applied voltage can accelerate or decelerate the baseline capillary-driven velocity, as well as be used to create a tunable valve that reversibly switches the flow on and off in an electrically controlled manner. The method relies on simple fabrication and allows repeated actuation, providing a high degree of flexibility for automation of liquid delivery. We adapt the Lucas-Washburn model to account for EO pumping and provide an experimentally validated analytical model for the distance penetrated by the liquid as a function of time and the applied voltage. We show that the EO-pump can reduce filling time by 6.5-fold for channels spanning several cm in length, relative to capillary filling alone. We demonstrate the utilization of the EO-pump for a tunable and dynamic flow control that accelerates, decelerates and stops the flow on demand. Finally, we present the use of the EO-pump for fluid flow sequencing on a paper-based device.

Extraction of electrokinetically separated analytes with on-demand encapsulation.

Microchip electrokinetic methods are capable of increasing the sensitivity of molecular assays by enriching and purifying target analytes. However, their use is currently limited to assays that can be performed under a high external electric field, as spatial separation and focusing is lost when the electric field is removed. We present a novel method that uses two-phase encapsulation to overcome this limitation. The method uses passive filling and pinning of an oil phase in hydrophobic channels to encapsulate electrokinetically separated and focused analytes with a brief pressure pulse. The resulting encapsulated sample droplet maintains its concentration over long periods of time without requiring an electric field and can be manipulated for further analysis, either on- or off-chip. We demonstrate the method by encapsulating DNA oligonucleotides in a 240 pL aqueous segment after isotachophoresis (ITP) focusing, and show that the concentration remains at 60% of the initial value for tens of minutes, a 22-fold increase over free diffusion after 20 minutes. Furthermore, we demonstrate manipulation of a single droplet by selectively encapsulating amplicon after ITP purification from a polymerase chain reaction (PCR) mix, and performing parallel off-chip detection reactions using the droplet. We provide geometrical design guidelines for devices implementing the encapsulation method, and show how the method can be scaled to multiple analyte zones.

Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics.

We present a novel method for real-time monitoring and kinetic analysis of fluorescence in situ hybridization (FISH). We implement the method using a vertical microfluidic probe containing a microstructure designed for rapid switching between probe solution and nonfluorescent imaging buffer. The FISH signal is monitored in real time during the imaging buffer wash, during which signal associated with unbound probes is removed. We provide a theoretical description of the method as well as a demonstration of its applicability using a model system of centromeric probes (Cen17). We demonstrate the applicability of the method for characterization of FISH kinetics under conditions of varying probe concentration, destabilizing agent (formamide) content, volume exclusion agent (dextran sulfate) content, and ionic strength. We show that our method can be used to investigate the effect of each of these variables and provide insight into processes affecting in situ hybridization, facilitating the design of new assays.

Nanoliter Cell Culture Array with Tunable Chemical Gradients.

A multitude of cell screening assays for diagnostic and research applications rely on quantitative measurements of a sample in the presence of different reagent concentrations. Standard methods rely on microtiter plates of varying well density, which provide simple and standardized sample addressability. However, testing hundreds of chemical dilutions requires complex automation, and typical well volumes of microtiter plates are incompatible with the analysis of a small number of cells. Here, we present a microfluidic device for creating a high-resolution chemical gradient spanning 200 nanoliter wells. Using air-based shearing, we show that the individual wells can be compartmentalized without altering the concentration gradient, resulting in a large set of isolated nanoliter cell culture wells. We provide an analytical and numerical model for predicting the concentration within each culture chamber and validate it against experimental results. We apply our system for the investigation of yeast cell metabolic gene regulation in the presence of different ratios of galactose/glucose concentrations and successfully resolve the nutrient threshold at which the cells activate the galactose pathway.