A Readily Scalable, Clinically Demonstrated, Antibiofouling Zwitterionic Surface Treatment for Implantable Medical Devices.

Unlike growth on tissue, microbes can grow freely on implantable devices with minimal immune system intervention and often form resilient biofilms that continuously pump out pathogenic cells. The efficacy of antibiotics used to treat infection is declining due to increased rates of pathogenic resistance. A simple, one-step zwitterionic surface modification is developed to significantly reduce protein and microbial adhesion to synthetic materials and demonstrate the successful modification of several clinically relevant materials, including recalcitrant materials such as elastomeric polydimethylsiloxane. The treated surfaces exhibit robust adhesion resistance against proteins and microorganisms in both static and flow conditions. Furthermore, the surface treatment prevents the adhesion of mammalian fibroblast cells while displaying no cytotoxicity. To demonstrate the clinical efficacy of the novel technology in the real-world, a surface-treated, commercial silicone foley catheter is developed that is cleared for use by the U.S. Food and Drug Administration (K192034). 16 long-term catheterized patients received surface-treated catheters and completed a Patient Global Impression of Improvement (PGI-I) questionnaire. 10 out of 16 patients described their urinary tract condition post implantation as "much better" or "very much better" and 72% (n = 13) of patients desire to continue using the surface-treated catheter over conventional latex or silicone catheters.

Monodisperse drops templated by 3D-structured microparticles.

The ability to create uniform subnanoliter compartments using microfluidic control has enabled new approaches for analysis of single cells and molecules. However, specialized instruments or expertise has been required, slowing the adoption of these cutting-edge applications. Here, we show that three dimensional-structured microparticles with sculpted surface chemistries template uniformly sized aqueous drops when simply mixed with two immiscible fluid phases. In contrast to traditional emulsions, particle-templated drops of a controlled volume occupy a minimum in the interfacial energy of the system, such that a stable monodisperse state results with simple and reproducible formation conditions. We describe techniques to manufacture microscale drop-carrier particles and show that emulsions created with these particles prevent molecular exchange, concentrating reactions within the drops, laying a foundation for sensitive compartmentalized molecular and cell-based assays with minimal instrumentation.

Fabrication of 3D concentric amphiphilic microparticles to form uniform nanoliter reaction volumes for amplified affinity assays.

Reactions performed in uniform microscale volumes have enabled numerous applications in the analysis of rare entities (e.g. cells and molecules). Here, highly monodisperse aqueous droplets are formed by simply mixing microscale multi-material particles, consisting of concentric hydrophobic outer and hydrophilic inner layers, with oil and water. The particles are manufactured in batch using a 3D printed device to co-flow four concentric streams of polymer precursors which are polymerized with UV light. The cross-sectional shapes of the particles are altered by microfluidic nozzle design in the 3D printed device. Once a particle encapsulates an aqueous volume, each "dropicle" provides uniform compartmentalization and customizable shape-coding for each sample volume to enable multiplexing of uniform reactions in a scalable manner. We implement an enzymatically-amplified immunoassay using the dropicle system, yielding a detection limit of <1 pM with a dynamic range of at least 3 orders of magnitude. Multiplexing using two types of shape-coded particles was demonstrated without cross talk, laying a foundation for democratized single-entity assays.

FlowSculpt: software for efficient design of inertial flow sculpting devices.

Flow sculpting is a powerful method for passive flow control that uses a sequence of bluff-body structures to engineer the structure of inertially flowing microfluidic streams. A variety of cross-sectional flow shapes can be created through this method, offering a new platform for flow manipulation or material fabrication useful in bioengineering, manufacturing, and chemistry applications. However, the inverse problem in flow sculpting - designing a device that produces a target fluid flow shape - remains challenging due to the complex, diverse, and enormous design space. Solutions to the inverse problem have been constrained to single-material fluid streams that are shaped into top-bottom symmetric shapes due to the bluff-body structures available in current libraries (pillars) that span the height of the channel. In this work, we introduce multi-material design and symmetry-breaking flow deformations enabled by half-height pillars, presented within an extremely fast simulation method for flow sculpting yielding a 34-fold reduction in runtime. The framework is deployed freely as a cross-platform application called "FlowSculpt". We detail its implementation and usage, and discuss the addition of enhanced search operations, which enable users to more easily design flow shapes that replicate their input drawings. With FlowSculpt, the microfluidics community can now quickly design flow shaping microfluidic devices on modest hardware, and easily integrate these complex physics into their research toolkit.

Shaped 3D microcarriers for adherent cell culture and analysis.

Standard tissue culture of adherent cells is known to poorly replicate physiology and often entails suspending cells in solution for analysis and sorting, which modulates protein expression and eliminates intercellular connections. To allow adherent culture and processing in flow, we present 3D-shaped hydrogel cell microcarriers, which are designed with a recessed nook in a first dimension to provide a tunable shear-stress shelter for cell growth, and a dumbbell shape in an orthogonal direction to allow for self-alignment in a confined flow, important for processing in flow and imaging flow cytometry. We designed a method to rapidly design, using the genetic algorithm, and manufacture the microcarriers at scale using a transient liquid molding optofluidic approach. The ability to precisely engineer the microcarriers solves fundamental challenges with shear-stress-induced cell damage during liquid-handling, and is poised to enable adherent cell culture, in-flow analysis, and sorting in a single format.

Rapid Software-Based Design and Optical Transient Liquid Molding of Microparticles.

Microparticles with complex 3D shape and composition are produced using a novel fabrication method, optical transient liquid molding, in which a 2D light pattern exposes a photopolymer precursor stream shaped along the flow axis by software-aided inertial flow engineering.

Research highlights: microfluidically-fabricated materials.

Polymer particles with precise shapes or chemistries are finding unique uses in a variety of applications, including tissue engineering, drug delivery, barcoding, and diagnostic imaging. Microfluidic systems have been and are continuing to play a large role in enabling the precision synthesis of designer particles in a uniform manner. To expand the impact of these microfluidic-fabricated materials additional fundamental capabilities should still be developed. The capability to fabricate microparticles with complex three-dimensional shapes and increase the production rate of particles to an industrial scale will allow evaluation of shaped particles in a range of new applications to enhance biological, magnetic, optical, surface wetting, as well as other interfacial or mechanical properties of materials. Here we highlight work applying large collections of simple spherical microgels, with unique surface chemistry that allows in situ particle-particle annealing, to form microporous injectable scaffolds for accelerated tissue regeneration. We also report on two other techniques that are addressing the ability to create 3D-shaped microparticles by first sculpting a fluid precursor stream, and increasing the rate of production of particles using contact lithography to millions of particles per hour. The combination of these capabilities and the applications they will enable suggest a bright future for microfluidics in making the next materials.

Micropillar sequence designs for fundamental inertial flow transformations.

The ability to control the shape of a flow in a passive microfluidic device enables potential applications in chemical reaction control, particle separation, and complex material fabrication. Recent work has demonstrated the concept of sculpting fluid streams in a microchannel using a set of pillars or other structures that individually deform a flow in a predictable pre-computed manner. These individual pillars are then placed in a defined sequence within the channel to yield the composition of the individual flow deformations - and ultimately complex user-defined flow shapes. In this way, an elegant mathematical operation can yield the final flow shape for a sequence without an experiment or additional numerical simulation. Although these approaches allow for programming complex flow shapes without understanding the detailed fluid mechanics, the design of an arbitrary flow shape of interest remains difficult, requiring significant design iteration. The development of intuitive basic operations (i.e. higher-level functions that consist of combinations of obstacles) that act on the flow field to create a basis for more complex transformations would be useful in systematically achieving a desired flow shape. Here, we show eight transformations that could serve as a partial basis for more complex transformations. We initially used in-house, freely available custom software (uFlow), which allowed us to arrive at these transformations that include making a fluid stream concave and convex, tilting, stretching, splitting, adding a vertex, shifting, and encapsulating another flow stream. The pillar sequences corresponding to these transformations were subsequently fabricated and optically analyzed using confocal imaging - yielding close agreement with uFlow-predicted shapes. We performed topological analysis on each transformation, characterizing potential sequences leading to these outputs and trends associated with changing diameter and placement of the pillars. We classify operations into four sets of sequence-building concatenations: stacking, recursion, mirroring, and shaping. The developed basis should help in the design of microfluidic systems that have a phenomenal variety of applications, such as optofluidic lensing, enhanced heat transfer, or new polymer fiber design.

Research highlights: increasing paper possibilities.

In this issue we highlight three recent papers that demonstrate new strategies to extend the capabilities of paper microfluidics. Paper (a mesh of porous fibers) has a long history as a substrate to perform biomolecular assays. Traditional lateral flow immunoassays (LFAs) are widely used for rapid diagnostic tests, and perform well when a yes or no answer is required and the analyte of interest is at relatively high concentrations. High concentrations are required because usually only a small volume of analyte-containing fluid flows past the detection region, leading to a limited signal. Further, the small pores within paper matrices prevent the use of paper to control the flow of larger particles and cells, limiting the use of paper microfluidics for cell-based diagnostics. The work we highlight addresses these important unmet challenges in paper microfluidics: enriching low concentration analytes to a higher concentration in a smaller volume that can be processed effectively, and using paper to pump flows in larger channels amenable to cells. Applying these new approaches may allow diagnosis of disease states currently technically unachievable using current LFA systems, while maintaining many of the "un-instrumented" advantages of an assay on self-wicking paper.

A microfluidic cell culture array with various oxygen tensions.

Oxygen tension plays an important role in regulating various cellular functions in both normal physiology and disease states. Therefore, drug testing using conventional in vitro cell models under normoxia often possesses limited prediction capability. A traditional method of setting an oxygen tension in a liquid medium is by saturating it with a gas mixture at the desired level of oxygen, which requires bulky gas cylinders, sophisticated control, and tedious interconnections. Moreover, only a single oxygen tension can be tested at the same time. In this paper, we develop a microfluidic cell culture array platform capable of performing cell culture and drug testing under various oxygen tensions simultaneously. The device is fabricated using an elastomeric material, polydimethylsiloxane (PDMS) and the well-developed multi-layer soft lithography (MSL) technique. The prototype device has 4 × 4 wells, arranged in the same dimensions as a conventional 96-well plate, for cell culture. The oxygen tensions are controlled by spatially confined oxygen scavenging chemical reactions underneath the wells using microfluidics. The platform takes advantage of microfluidic phenomena while exhibiting the combinatorial diversities achieved by microarrays. Importantly, the platform is compatible with existing cell incubators and high-throughput instruments (liquid handling systems and plate readers) for cost-effective setup and straightforward operation. Utilizing the developed platform, we successfully perform drug testing using an anti-cancer drug, triapazamine (TPZ), on adenocarcinomic human alveolar basal epithelial cell line (A549) under three oxygen tensions ranging from 1.4% to normoxia. The developed platform is promising to provide a more meaningful in vitro cell model for various biomedical applications while maintaining desired high throughput capabilities.

Electrofluidic pressure sensor embedded microfluidic device: a study of endothelial cells under hydrostatic pressure and shear stress combinations.

Various microfluidic cell culture devices have been developed for in vitro cell studies because of their capabilities to reconstitute in vivo microenvironments. However, controlling flows in microfluidic devices is not straightforward due to the wide varieties of fluidic properties of biological samples. Currently, flow observations mainly depend on optical imaging and macro scale transducers, which usually require sophisticated instrumentation and are difficult to scale up. Without real time monitoring, the control of flows can only rely on theoretical calculations and numerical simulations. Consequently, these devices have difficulty in being broadly exploited in biological research. This paper reports a microfluidic device with embedded pressure sensors constructed using electrofluidic circuits, which are electrical circuits built by fluidic channels filled with ionic liquid. A microfluidic device culturing endothelial cells under various shear stress and hydrostatic pressure combinations is developed to demonstrate this concept. The device combines the concepts of electrofluidic circuits for pressure sensing, and an equivalent circuit model to design the cell culture channels. In the experiments, human umbilical vein endothelial cells (HUVECs) are cultured in the device with a continuous medium perfusion, which provides the combinatory mechanical stimulations, while the hydrostatic pressures are monitored in real time to ensure the desired culture conditions. The experimental results demonstrate the importance of real time pressure monitoring, and how both mechanical stimulations affect the HUVEC culture. This developed microfluidic device is simple, robust, and can be easily scaled up for high-throughput experiments. Furthermore, the device provides a practical platform for an in vitro cell culture under well-controlled and dynamic microenvironments.

Integrated electrofluidic circuits: pressure sensing with analog and digital operation functionalities for microfluidics.

Microfluidic technology plays an essential role in various lab on a chip devices due to its desired advantages. An automated microfluidic system integrated with actuators and sensors can further achieve better controllability. A number of microfluidic actuation schemes have been well developed. In contrast, most of the existing sensing methods still heavily rely on optical observations and external transducers, which have drawbacks including: costly instrumentation, professional operation, tedious interfacing, and difficulties of scaling up and further signal processing. This paper reports the concept of electrofluidic circuits - electrical circuits which are constructed using ionic liquid (IL)-filled fluidic channels. The developed electrofluidic circuits can be fabricated using a well-developed multi-layer soft lithography (MSL) process with polydimethylsiloxane (PDMS) microfluidic channels. Electrofluidic circuits allow seamless integration of pressure sensors with analog and digital operation functions into microfluidic systems and provide electrical readouts for further signal processing. In the experiments, the analog operation device is constructed based on electrofluidic Wheatstone bridge circuits with electrical outputs of the addition and subtraction results of the applied pressures. The digital operation (AND, OR, and XOR) devices are constructed using the electrofluidic pressure controlled switches, and output electrical signals of digital operations of the applied pressures. The experimental results demonstrate the designed functions for analog and digital operations of applied pressures are successfully achieved using the developed electrofluidic circuits, making them promising to develop integrated microfluidic systems with capabilities of precise pressure monitoring and further feedback control for advanced lab on a chip applications.

Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions.

This paper reports a microfluidic device capable of generating oxygen gradients for cell culture using spatially confined chemical reactions with minimal chemical consumption. The microfluidic cell culture device is constructed by single-layer polydimethylsiloxane (PDMS) microfluidic channels, in which the cells can be easily observed by microscopes. The device can control the oxygen gradients without the utilization of bulky pressurized gas cylinders, direct addition of oxygen scavenging agents, or tedious gas interconnections and sophisticated flow control. In addition, due to the efficient transportation of oxygen within the device using the spatially confined chemical reactions, the microfluidic cell culture device can be directly used in conventional cell incubators without altering their gaseous compositions. The oxygen gradients generated in the device are numerically simulated and experimentally characterized using an oxygen-sensitive fluorescence dye. In this paper, carcinomic human alveolar basal epithelial (A549) cells have been cultured in the microfluidic device with a growth medium and an anti-cancer drug (Tirapazamine, TPZ) under various oxygen gradients. The cell experiment results successfully demonstrate the hyperoxia-induced cell death and hypoxia-induced cytotoxicity of TPZ. In addition, the results confirm the great cell compatibility and stable oxygen gradient generation of the developed device. Consequently, the microfluidic cell culture device developed in this paper is promising to be exploited in biological labs with minimal instrumentation to study cellular responses under various oxygen gradients.

Integrated ionic liquid-based electrofluidic circuits for pressure sensing within polydimethylsiloxane microfluidic systems.

This paper reports a novel pressure sensor with an electrical readout based on electrofluidic circuits constructed by ionic liquid (IL)-filled microfluidic channels. The developed pressure sensor can be seamlessly fabricated into polydimethylsiloxane (PDMS) microfluidic systems using the well-developed multilayer soft lithography (MSL) technique without additional assembly or sophisticated cleanroom microfabrication processes. Therefore, the device can be easily scaled up and is fully disposable. The pressure sensing is achieved by measuring the pressure-induced electrical resistance variation of the constructed electrofluidic resistor. In addition, an electrofluidic Wheatstone bridge circuit is designed for accurate and stable resistance measurements. The pressure sensor is characterized using pressurized nitrogen gas and various liquids which flow into the microfluidic channels. The experimental results demonstrate the great long-term stability (more than a week), temperature stability (up to 100 °C), and linear characteristics of the developed pressure sensing scheme. Consequently, the integrated microfluidic pressure sensor developed in this paper is promising for better monitoring and for characterizing the flow conditions and liquid properties inside the PDMS microfluidic systems in an easier manner for various lab on a chip applications.