Fluidic Network Research Articles

Cell-Free Gene Expression in Bioprinted Fluidic Networks.

The realization of soft robotic devices with life-like properties requires the engineering of smart, active materials that can respond to environmental cues in similar ways as living cells or organisms. Cell-free expression systems provide an approach for embedding dynamic molecular control into such materials that avoids many of the complexities associated with genuinely living systems. Here, we present a strategy to integrate cell-free protein synthesis within agarose-based hydrogels that can be spatially organized and supplied by a synthetic vasculature. We first utilize an indirect printing approach with a commercial bioprinter and Pluronic F-127 as a fugitive ink to define fluidic channel structures within the hydrogels. We then investigate the impact of the gel matrix on the expression of proteins in E. coli cell-extract, which is found to depend on the gel density and the dilution of the expression system. When supplying the vascularized hydrogels with reactants, larger components such as DNA plasmids are confined to the channels or immobilized in the gels while nanoscale reaction components can diffusively spread within the gel. Using a single supply channel, we demonstrate different spatial protein concentration profiles emerging from different cell-free gene circuits comprising production, gene activation, and negative feedback. Variation of the channel design allows the creation of specific concentration profiles such as a long-term stable gradient or the homogeneous supply of a hydrogel with proteins.

Geometrically Scalable Iontronic Memristors: Employing Bipolar Polyelectrolyte Gels for Neuromorphic Systems.

Iontronics that are capable of mimicking the functionality of biological systems within an artificial fluidic network have long been pursued for biomedical applications and ion-based intelligence systems. Here, we report on facile and robust realization of iontronic bipolar memristors featuring a three-layer polyelectrolyte gel structure. Significant memristive hysteresis of ion currents was successfully accomplished, and the memory time proved geometrically scalable from 200 to 4000 s. These characteristics were enabled by the ion concentration polarization-induced rectification ratio within the polyelectrolyte gels. The memristors exhibited memory dynamics akin to those observed in unipolar devices, while the bipolar structure notably enabled prolonged memory time and enhanced the ion conductance switching ratio with mesoscale (10-1000 μm) geometry precision. These properties endow the devices with the capability of effective neuromorphic processing with pulse-based input voltage signals. Owing to their simple fabrication process and superior memristive performance, the presented iontronic bipolar memristors are versatile and can be easily integrated into small-scale iontronic circuits, thereby facilitating advanced neuromorphic computing functionalities.

Phosphatidylinositol-4,5-biphosphate (PIP2)-Dependent Thermoring Basis for Cold-Sensing of the Transient Receptor Potential Melastatin-8 (TRPM8) Biothermometer

The menthol sensor transient receptor potential melastatin-8 (TRPM8) can be activated by cold and, thus, serves as a biothermometer in a primary afferent sensory neuron for innocuous-to-noxious cold detection. However, the precise structural origins of specific temperature thresholds and sensitivity have remained elusive. Here, a grid thermodynamic model was employed, to examine if the temperature-dependent noncovalent interactions found in the 3-dimensional (3D) structures of thermo-gated TRPM8 could assemble into a well-organized fluidic grid-like mesh network, featuring the constrained grids as the thermorings for cold-sensing in response to PIP2, Ca2+ and chemical agents. The results showed that the different interactions of TRPM8 with PIP2 during the thermal incubation induced the formation of the biggest grids with distinct melting temperature threshold ranges. Further, the overlapped threshold ranges between open and pre-open closed states were required for initial cold activation with the matched thermo-sensitivity and the decrease in the systematic thermal instability. Finally, the intact anchor grid near the lower gate was important for channel opening with the active selectivity filter. Thus, PIP2-dependent thermorings in TRPM8 may play a pivotal role in cold sensing.

Scalability of nanopore osmotic energy conversion

AbstractArtificial nanofluidic networks are emerging systems for blue energy conversion that leverages surface charge‐derived permselectivity to induce voltage from diffusive ion transport under salinity difference. Here the pivotal significance of electrostatic inter‐channel couplings in multi‐nanopore membranes, which impose constraints on porosity and subsequently influence the generation of large osmotic power outputs, is illustrated. Constructive interference is observed between two 20 nm nanopores of 30 nm spacing that renders enhanced permselectivity to osmotic power output via the recovered electroneutrality. On contrary, the interference is revealed as destructive in two‐dimensional arrays causing significant deteriorations of the ion selectivity even for the nanopores sparsely distributed at an order of magnitude larger spacing than the Dukhin length. Most importantly, a scaling law is provided for deducing the maximal membrane area and porosity to avoid the selectivity loss via the inter‐pore electrostatic coupling. As the electric crosstalk is inevitable in any fluidic network, the present findings can be a useful guide to design nanoporous membranes for scalable osmotic power generations.

Multifunctional droplet handling on surface-charge-graphic-decorated porous papers.

Developing a fluidic platform that combines high-throughput with reconfigurability is essential for a wide range of cutting-edge applications, but achieving both capabilities simultaneously remains a significant challenge. Herein, we propose a novel and unique method for droplet manipulation via drawing surface charge graphics on electrode-free papers in a contactless way. We find that opposite charge graphics can be written and retained on the surface layer of porous insulating paper by a controlled charge depositing method. The retained charge graphics result in high-resolution patterning of electrostatic potential wells (EPWs) on the hydrophobic porous surface, allowing for digital and high-throughput droplet handling. Since the charge graphics can be written/projected dynamically and simultaneously in large areas, allowing for on-demand and real-time reconfiguration of EPWs, we are able to develop a charge-graphic fluidic platform with both high reconfigurability and high throughput. The advantages and application potential of the platform have been demonstrated in chemical detection and dynamically controllable fluidic networks.

Flow tweezing of anisotropic magnetic microrobots in a dynamic magnetic trap for active retention and localized flow sensing.

Controlled manipulation of microscale robotic devices in complex fluidic networks is critical for various applications in biomedical endovascular sensing, lab-on-chip biochemical assays, and environmental monitoring. However, achieving controlled transport and active retention of microscale robots with flow sensing capability has proven to be challenging. Here, we report the dynamic tweezing of an anisotropic magnetic microrobot in a rotating magnetic trap for active retention and localized flow sensing under confined fluidic conditions. We reveal a series of unconventional motion modes and the dynamics of the microrobot transporting in a confined fluidic flow, which manifest themselves as transitions from on-trap centre rolling to large-area revolution and off-trap centre rolling with varying rotating frequencies. By retaining the robot within the magnetic trap and its motion modulated by the field frequency, the off-centre rolling of the microrobot endows it with crucial localized flow sensing capabilities, including flow rate and flow direction determination. The magnetic microrobot serves as a mobile platform for measuring the flow profile along a curved channel, mimicking a blood vessel. Our findings unlock a new strategy to determine the local magnetic tweezing force profile and flow conditions in arbitrary flow channels, revealing strong potential for microfluidics, chemical reactors, and in vivo endovascular flow measurement.

A biphasic accelerated strand exchange amplification strategy for culture-independent and rapid detection of Salmonella enterica in food samples.

Salmonella enterica is a common foodborne pathogen that can cause food poisoning in humans. The organism also infects and causes disease in animals. Rapid and sensitive detection of S. enterica is essential to prevent the spread of this pathogen. Traditional technologies for the extraction and detection of this pathogen from complex food matrices are cumbersome and time-consuming. In this study, we introduced a novel strategy of biphasic assay integrated with an accelerated strand exchange amplification (ASEA) method for efficient detection of S. enterica without culture or other extraction procedures. Food samples are rapidly dried, resulting in a physical fluidic network inside the dried food matrix, which allows polymerases and primers to access the target DNA and initiate ASEA. The dried food matrix is defined as the solid phase, while amplification products are enriched in the supernatant (liquid phase) and generate fluorescence signals. The analytical performances demonstrated that this strategy was able to specifically identify S. enterica and did not show any cross-reaction with other common foodborne pathogens. For artificially spiked food samples, the strategy can detect 5.0 × 101 CFU mL-1S. enterica in milk, 1.0 × 102 CFU g-1 in duck, scallop or lettuce, and 1.0 × 103 CFU g-1 in either oyster or cucumber samples without pre-enrichment of the target pathogen. We further validated the strategy using 82 real food samples, and this strategy showed 92% sensitivity. The entire detection process can be finished, sample-to-answer, within 50 min, dramatically decreasing the detection time. Therefore, we believe that the proposed method enables rapid and sensitive detection of S. enterica and holds great promise for the food safety industry.

Programmable fluidic networks on centrifugal microfluidic discs

BackgroundBiomedical diagnostic and lab automation solutions built on the Lab-on-a-Disc (LoaD) platform has great potential due to their independence from specialised micro-pumps and their ease of integration, through direct pipetting, with manual or automated workflows. However, a challenge for all microfluidic chips is their cost of manufacture when each microfluidic disc must be customized for a specific application. In this paper, we present centrifugal discs with programmable fluidic networks. ResultsBased on dissolvable film valves, we present two technologies. The first, based on recently introduced pulse-actuated dissolvable film valves, is a centrifugal disc which, depending on how it is loaded, is configured to perform either six sequential reagent releases through one reaction chamber or three sequential reagent releases through two reaction chambers. In the second approach, we use the previously introduced electronic Lab-on-a-Disc (eLoaD) wireless valve array, which can actuate up to 128 centrifugo-pneumatic dissolvable film valves in a pre-defined sequence. In this approach we present a disc which can deliver any one of 8 reagent washes to any one of four reaction chambers. We use identical discs to demonstrate the first four sequential washes through two reaction chambers and then two sequential washes through four reaction chambers. SignificanceThese programmable fluidic networks have the potential to allow a single disc architecture to be applied to multiple different assay types and so can offer a lower-cost and more integrated alternative to the standard combination of micro-titre plate and liquid handling robot. Indeed, it may even be possible to conduct multiple different assays concurrently. This can have the effect of reducing manufacturing costs and streamlining supply-chains and so results in a more accessible diagnostic platform.

Thermoring basis for the TRPV3 bio-thermometer

The thermosensitive transient receptor potential (TRP) channels are well-known as bio-thermometers with specific temperature thresholds and sensitivity. However, their precise structural origins are still mysterious. Here, graph theory was used to test how the temperature-dependent non-covalent interactions as identified in the 3D structures of thermo-gated TRPV3 could form a systematic fluidic grid-like mesh network with the constrained thermo-rings from the biggest grids to the smallest ones as necessary structural motifs for the variable temperature thresholds and sensitivity. The results showed that the heat-evoked melting of the biggest grids may control the specific temperature thresholds to initiate channel gating while the smaller grids may be required to secure heat efficacy. Together, all the grids along the lipid-dependent minimal gating pathway may be necessary to change with molar heat capacity for the specific temperature sensitivity. Therefore, this graph theory-based grid thermodynamic model may provide an extensive structural basis for the thermo-gated TRP channels.

Theoretical analysis of immunochromatographic assay and consideration of its operating parameters for efficient designing of high-sensitivity cardiac troponin I (hs-cTnI) detection

Troponin is the American College of Cardiology and American Heart Association preferred biomarker for diagnosing acute myocardial infarction (MI). We provide a modeling framework for high sensitivity cardiac Troponin I (hs-cTnI) detection in chromatographic immunoassays (flow displacement mode) with an analytical limit of detection, i.e., LOD < 10 ng/L. We show that each of the various control parameters exert a significant influence over the design requirements to reach the desired LOD. Additionally, the design implications in a multiplexed fluidic network, as in the case of Simple Plex™ Ella instrument, are significantly affected by the choice of the number of channels or partitions in the network. We also provide an upgrade on the existing LOD equation to evaluate the necessary minimum volume to detect a particular concentration by considering the effects of stochastics and directly incorporating the target number of copies in each of the partitions in case of multiplexed networks. Even though a special case of cTnI has been considered in this study, the model and analysis are analyte agnostic and may be applied to a wide class of chromatographic immunoassays. We believe that this contribution will lead to more efficient designing of the immunochromatographic assays.

Memory-induced Magnus effect

Spinning objects moving through air or a liquid experience a lift force—a phenomenon known as the Magnus effect. This effect is commonly exploited in ball sports but also is of considerable importance for applications in the aviation industry. Whereas Magnus forces are strong for large objects, they are weak at small scales and eventually vanish for overdamped micrometre-sized particles in simple liquids. Here we demonstrate a roughly one-million-fold enhanced Magnus force of spinning colloids in viscoelastic fluids. Such fluids are characterized by a time-delayed response to external perturbations, which causes a deformation of the fluidic network around the moving particle. When the particle also spins, the deformation field becomes misaligned relative to the particle’s moving direction, leading to a force perpendicular to the direction of travel and the spinning axis. Our uncovering of strongly enhanced memory-induced Magnus forces at microscales opens up applications for particle sorting and steering, and the creation and visualization of anomalous flows.

Spatiotemporal Pattern Formation of Metal Electrodissolution Reactions Using Multielectrode Arrays in Lab-on-Chip Cells

Electrochemical reactions can generate complex spatiotemporal patterns due to the inherent nonlinearities in the reaction kinetics, which often couple with various mass transfer and electrical coupling effects. We explore spatiotemporal pattern formation using multielectrode arrays embedded in flow channels. The kinetic equations are expressed as a superposition of local kinetics with a network of electrical coupling due to ohmic potential drops in the electrolyte. The networks are revealed by analysis of the synchronization patterns with the use of an oscillatory chemical reaction (nickel electrodissolution) and are further confirmed by direct decoding using phase model analysis. In dual electrode configuration, a variety coupling schemes, (uni- or bidirectional positive or negative) were identified depending on the relative placement of the reference and counter electrodes (e.g., placed at the same or the opposite ends of the ow channel). The synchronization patterns and the network configuration are compared for the linear and the branched channel configurations. With straight channel and multiple electrodes, a scale-free network was formed. With branched channels, the global to local coupling can be changed by the position of electrodes in the channel, and introduction of different concentration of electrolyte can mediate coupling between only the downstream electrodes. In fluidic network with cross channels, nonlinearity can be induced by obstacles in the channel which open ways to investigate the Braess’s paradox. The networked, electrode potential (current) spike generating electrochemical reactions hold potential for construction of an in-situ information processing unit to be used in electrochemical devices in sensors and batteries.

Integrative modeling of hemodynamic changes and perfusion impairment in coronary microvascular disease.

Introduction: Coronary microvascular disease is one of the responsible factors for cardiac perfusion impairment. Due to diagnostic and treatment challenges, this pathology (characterized by alterations to microvasculature local hemodynamics) represents a significant yet unsolved clinical problem. Methods: Due to the poor understanding of the onset and progression of this disease, we propose a new and noninvasive strategy to quantify in-vivo hemodynamic changes occurring in the microvasculature. Specifically, we here present a conceptual workflow that combines both in-vitro and in-silico modelling for the analysis of the hemodynamic alterations in the microvasculature. Results: First, we demonstrate a hybrid additive manufacturing process to fabricate circular cross-section, biocompatible fluidic networks in polytetrafluoroethylene. We then use these microfluidic devices and computational fluid dynamics to simulate different degrees of perfusion impairment. Discussion: Ultimately, we show that the developed workflow defines a robust platform for the multiscale analysis of multifactorial events occurring in coronary microvascular disease.

Thermoring-based heat activation switches in the TRPV1 biothermometer

Non-covalent interactions in bio-macromolecules are individually weak but collectively important. How they take a concerted action in a complex biochemical reaction network to realize their thermal stability and activity is still challenging to study. Here graph theory was used to investigate how the temperature-dependent non-covalent interactions as identified in the 3D structures of the thermo-gated capsaicin receptor TRPV1 could form a systemic fluidic grid-like mesh network with topological grids constrained as the thermo-rings to govern heat-sensing. The results showed that the heat-evoked melting of the biggest grid initiated a matched temperature threshold to release the lipid from the active vanilloid site for channel activation. Meanwhile, smaller grids were required to stabilize heat efficacy. Altogether, the change in the total grid sizes upon the change in the total noncovalent interactions along the lipid-dependent gating pathway was necessary for the matched temperature sensitivity. Therefore, this grid thermodynamic model may be broadly significant for the structural thermostability and the functional thermoactivity of bio-macromolecules.

Engineering of branched fluidic networks that minimise energy dissipation

Power minimisation of fluid transport in branched fluidic networks has become of paramount importance for microfluidics, additive manufacturing and hierarchical functional materials. For fully developed laminar flow of Newtonian fluids, Murray's theory provides a solution for the channel and network dimensions that minimise power consumption. However, design and optimisation of networks that transport complex fluids is still challenging. Here, we generalise Murray's theory towards fluid rheologies, including non-Newtonian (power-law) and yield-stress fluids (Bingham, Herschel–Bulkley, Casson). A straightforward graphical approach is presented that provides the optimal radii in a branching network, and the angles between these branches. The wall shear stress is found to be uniform over the entire network, and the velocity profile is self-similar. Furthermore, the effect of non-optimal channel radii on the power consumption of the network is investigated. Finally, examples illustrate how this approach applies to a wide variety of systems.

Thermal Ring-Based Heat Switches in Hyperthermophilic Class II Bacterial Fructose Aldolase.

Both thermophilic and hyperthermophilic enzymes in bacterial and archaeal species are activated above a specific temperature threshold but inactivated at another higher temperature. However, the underlying structural basis for these two heat switches is still unresolved. Here, graph theory was used to test if the temperature-dependent noncovalent interactions and metal bridges as identified in a series of crystal structures of the class II bacterial fructose 1,6-bisphosphate aldolase homodimer or homotetramer with or without natural substrates and products bound could form systematic fluidic grid-like mesh networks with topological grids as thermal rings to regulate their structural thermostability and functional thermoactivity. The results indicated that the second biggest grid in the Thermus aquaticus fructose 1,6-diphosphate aldolase dimer may control the specific temperature threshold to release the swapping flexible active sites at the dimeric interface for heat-evoked activation. Meanwhile, the third biggest grid may serve as a necessary structural motif against heat inactivation. Finally, the smallest grid may act as a stiff thermostable anchor. Its dissociation at the maximal melting temperature threshold may stop the catalytic activity. Taken as a whole, this computational study may render the structural motifs for the optimal growth temperature and the extreme heat stability of hyperthermophiles.

Fluid Flow Control in Cotton Threads with Mesoporous Silica Coatings

AbstractMicrofluidic devices are important, e.g. in the field of point of care diagnostics. They are of special importance, if they are fabricated out of cheap and renewable materials. Tackling complex separation or sensing problems profits from modular three‐dimensional fluidic devices. Using cotton threads as renewable material allows the modular design of three‐dimensional fluidic devices and networks. Here, fluidic threads with modular designed and tunable thread wettability are presented. The wettability is gradually adjusted from highly hydrophilic to hydrophobic. The thread wettability directly affects the fluid imbibition velocity as well as the distance, which the fluid imbibes into the thread. The wettability adjustment is based on a simple dense or mesoporous silica coating applied onto the cotton thread using sol‐gel chemistry and evaporation induced self‐assembly. In addition to wettability, the mesoporosity and the pore functionalization are used to tune the fluid velocity within the thread. Connecting different silica functionalized threads into one device by knotting them together, fluids can be guided through this network in a predicted manner, which allows a modular design of 3D microfluidic thread‐based devices.

Network Basis for the Heat-Adapted Structural Thermostability of Bacterial Class II Fructose Bisphosphate Aldolase.

The sufficient structural thermostability of a biological macromolecule is an overriding need for green nanoreactors and nanofactories to secure high activity. However, little is still known about what specific structural motif is responsible for it. Here, graph theory was employed to examine if the temperature-dependent noncovalent interactions and metal bridges, as identified in the structures of Escherichia coli class II fructose 1,6-bisphosphate aldolase, could shape a systematic fluidic grid-like mesh network with topological grids to regulate the structural thermostability of the wild-type construct and its evolved variants in each generation upon decyclization. The results indicated that the biggest grids may govern the temperature thresholds for their tertiary structural perturbations but without affecting the catalytic activities. Moreover, lower grid-based systematic thermal instability may facilitate structural thermostability, but a highly independent thermostable grid may still be required to serve as a critical anchor to secure the stereospecific thermoactivity. Its end melting temperature thresholds, together with the start ones of the biggest grids in the evolved variants, may confer high temperature sensitivity against thermal inactivation. Collectively, this computational study may have widespread significance in advancing our complete understanding and biotechnology of the thermoadaptive mechanism of the structural thermostability of a biological macromolecule.

The Network Basis for the Structural Thermostability and the Functional Thermoactivity of Aldolase B

Thermostability is important for the thermoactivity of proteins including enzymes. However, it is still challenging to pinpoint the specific structural factors for different temperature thresholds to initiate their specific structural and functional perturbations. Here, graph theory was used to investigate how the temperature-dependent noncovalent interactions as identified in the structures of aldolase B and its prevalent A149P mutant could form a systematic fluidic grid-like mesh network with topological grids to regulate the structural thermostability and the functional thermoactivity upon cyclization against decyclization in an extended range of a subunit. The results showed that the biggest grid may determine the melting temperature thresholds for the changes in their secondary and tertiary structures and specific catalytic activities. Further, a highly conserved thermostable grid may serve as an anchor to secure the flexible active site to achieve the specific thermoactivity. Finally, higher grid-based systematic thermal instability may disfavor the thermoactivity. Thus, this computational study may provide critical clues for the structural thermostability and the functional thermoactivity of proteins including enzymes.

Integrating Bio-Sensing Array with Blood Plasma Separation on a Centrifugal Platform.

Numerous immunoassays have been successfully integrated on disc-based centrifugal platforms (CDs) over the last 20 years. These CD devices can be used as portable point-of-care (POC) platforms with sample-to-answer capabilities where bodily fluids such as whole blood can be used as samples directly without pre-processing. In order to use whole blood as a sample on CDs, centrifugation is used to separate red blood cells from plasma on CDs. There are several techniques for using specific fluidic patterns in the centrifugal fluidic network, such as reciprocation, that enhances the sensitivity of the immunoassays, including those using microarray antigen membranes. Present work demonstrates, for the first time, simultaneous integration of blood plasma separation (BPS) and reciprocation on the CD platform. The integrated design allows plasma that is separated from the red blood cells in a sedimentation chamber to flow into the reciprocation chamber via a narrow connecting channel of 0.5 mm × 0.5 mm cross-section. Due to the small cross-section of the connecting channel, there is no inflow of the red blood cell into the reciprocation chamber during subsequent fluidic operations of the CD. While no inflow of the red blood cells into the reciprocation chamber was observed, the conditions of 20 g jerk acceleration were also simulated in ANSYS finite element analysis software, and it was found that the CD design that was used is capable of retaining red blood cells in the sedimentation chamber. Experimentally, the isolation of red blood cells in the sedimentation chamber was confirmed using the ImageJ image processor to detect the visible color-based separation of the plasma from the blood. A fluorescent analyte testing on the bio-sensing array of the presented novel integrated design and on the standard reciprocation design CD was conducted for 7 min of reciprocation in each case. The test analyte was Europium Streptavidin Polystyrene analyte (10-3 mg/mL) and the microarray consisted of Biotin bovine serum albumin (BSA) dots. The fluorescent signals for the standard and integrated designs were nearly identical (within the margin of error) for the first several minutes of reciprocation, but the fluorescent signal for the integrated design was significantly higher when the reciprocation time was increased to 7 min.