Autonomous capillary microfluidic devices with constant flow rate and temperature-controlled valving.

In this paper, we report on a capillary microfluidic device with constant flow rate and temperature-triggered stop valve function. It contains a PDMS channel that was grafted by a thermo-responsive polymer poly(N-isopropylacrylamide) (PNIPAm). The channel exhibits a constant capillary filling speed. By locally increasing the temperature in the channel from 20 °C to 37 °C using a microfabricated heater, a change of the surface wettability from hydrophilic to hydrophobic is obtained creating a hydrophobic stop valve. The valve can be reopened by lowering the temperature. The device is simple to fabricate and can be used as an actuatable capillary pump operating around room temperature. To understand the constant capillary filling speed, we performed contact angle measurements, in which we found slow wetting kinetics of PNIPAm-g-PDMS surfaces at temperatures below the lower critical solution temperature (LCST) of PNIPAm and fast wetting kinetics above the LCST. We interpret this as the result of the diffusive hydration process of PNIPAm below the LCST and the absence of hydration on the hydrophobic PNIPAm thin layer above the LCST.

Giant and explosive plasmonic bubbles by delayed nucleation.

When illuminated by a laser, plasmonic nanoparticles immersed in water can very quickly and strongly heat up, leading to the nucleation of so-called plasmonic vapor bubbles. While the long-time behavior of such bubbles has been well-studied, here, using ultrahigh-speed imaging, we reveal the nucleation and early life phase of these bubbles. After some delay time from the beginning of the illumination, a giant bubble explosively grows, and collapses again within 200 µs (bubble life phase 1). The maximal bubble volume [Formula: see text] remarkably increases with decreasing laser power, leading to less total dumped energy E. This dumped energy shows a universal linear scaling relation with [Formula: see text], irrespective of the gas concentration of the surrounding water. This finding supports that the initial giant bubble is a pure vapor bubble. In contrast, the delay time does depend on the gas concentration of the water, as gas pockets in the water facilitate an earlier vapor bubble nucleation, which leads to smaller delay times and lower bubble nucleation temperatures. After the collapse of the initial giant bubbles, first, much smaller oscillating bubbles form out of the remaining gas nuclei (bubble life phase 2). Subsequently, the known vaporization dominated growth phase takes over, and the bubble stabilizes (life phase 3). In the final life phase 4, the bubble slowly grows by gas expelling due to heating of the surrounding. Our findings on the explosive growth and collapse during the early life phase of a plasmonic vapor bubble have strong bearings on possible applications of such bubbles.

Free Flow Ion Concentration Polarization Focusing (FF-ICPF).

Electrokinetic separation techniques in microfluidics are a powerful analytical chemistry tool, although an inherent limitation of microfluidics is their low sample throughput. In this article we report a free-flow variant of an electrokinetic focusing method, namely ion concentration polarization focusing (ICPF). The analytes flow continuously through the system via pressure driven flow while they separate and concentrate perpendicularly to the flow by ICPF. We demonstrate free flow ion concentration polarization focusing (FF-ICPF) in two operating modes, namely peak and plateau modes. Additionally, we showed the separation resolution could be improved by the use of an electrophoretic spacer. We report a concentration factor of 10 in human blood plasma in continuous flow at a flow rate of 15 µL min-1.

Mass Transport Determined Silica Nanowires Growth on Spherical Photonic Crystals with Nanostructure-Enabled Functionalities.

A robust and facile method has been developed to obtain directional growth of silica nanowires (SiO2 NWs) by regulating mass transport of silicon monoxide (SiO) vapor. SiO2 NWs are grown by vapor-liquid-solid (VLS) process on a surface of gold-covered spherical photonic crystals (SPCs) annealed at high temperature in an inert gas atmosphere in the vicinity of a SiO source. The SPCs are prepared from droplet confined colloidal self-assembly. SiO2 NW morphology is governed by diffusion-reaction process of SiO vapor, whereby directional growth of SiO2 NWs toward the low SiO concentration is obtained at locations with a high SiO concentration gradient, while random growth is observed at locations with a low SiO concentration gradient. Growth of NWs parallel to the supporting substrate surface is of great importance for various applications, and this is the first demonstration of surface-parallel growth by controlling mass transport. This controllable NW morphology enables production of SPCs covered with a large number of NWs, showing multilevel micro-nano feature and high specific surface area for potential applications in superwetting surfaces, oil/water separation, microreactors, and scaffolds. In addition, the controllable photonic stop band properties of this hybrid structure of SPCs enable the potential applications in photocatalysis, sensing, and light harvesting.

Integrated internal standards: A sample prep-free method for better precision in microchip CE.

Point-of-care systems based on microchip capillary electrophoresis require single-use, disposable microchips prefilled with all necessary solutions so an untrained operator only needs to apply the sample and perform the analysis. While microchip fabrication can be (and has been) standardized, some manufacturing differences between microchips are unavoidable. To improve analyte precision without increasing device costs or introducing additional error sources, we recently proposed the use of integrated internal standards (ISTDs): ions added to the BGE in small concentrations which form system peaks in the electropherogram that can be used as a measurement reference. Here, we further expand this initial proof-of-principle test to study a clinically-relevant application of K ion concentrations in human blood; however, using a mock blood solution instead of real samples to avoid interference from other obstacles (e.g. cell lysis). Cs as an integrated ISTD improves repeatability of K ion migration times from 6.97% to 0.89% and the linear calibration correlation coefficient (R2 ) for K quantification from 0.851 to 0.967. Peak area repeatability improves from 11.6-13.3% to 4.75-5.04% at each K concentration above the LOQ. These results further validate the feasibility of using integrated ISTDs to improve imprecision in disposable microchip CE devices by demonstrating their application for physiological samples.

Ion Concentration Polarization for Microparticle Mesoporosity Differentiation.

Microparticle porosity is normally determined in bulk manner providing an ensemble average that hinders establishing the individual role of each microparticle. On the other hand, single particle characterization implies expensive technology. We propose to use ion concentration polarization to measure differences in mesoporosity at the single particle level. Ion concentration polarization occurs at the interface between an electrolyte and a porous particle when an electric field is applied. The extent of ion concentration polarization depends, among others, on the mesopore size and density. By using a fluorescence marker, we could measure differences in concentration polarization between particles with 3 and 13 nm average mesopore diameters. A qualitative model was developed in order to understand and interpret the phenomena. We believe that this inexpensive method could be used to measure differences in mesoporous particle materials such as catalysts.

Engulfment control of platinum nanoparticles into oxidized silicon substrates for fabrication of dense solid-state nanopore arrays.

We found that platinum (Pt) nanoparticles, upon annealing at high temperature of 1000 °C, are engulfed into amorphous fused-silica or thermal oxide silicon substrates. The same phenomenon was previously published for gold (Au) nanoparticles. Similar to the Au nanoparticles, the engulfed Pt nanoparticles connect to the surface of the substrates through conical nanopores, and the size of the Pt nanoparticles decreases with increasing depth of the nanopores. We explain the phenomena as driven by the formation of platinum oxide by reaction of the platinum with atmospheric oxygen, with platinum oxide evaporating to the environment. We found that the use of Pt provides much better controllability than the use of Au. Due to the high vapor pressure of platinum oxide, the engulfment of the Pt nanoparticles into oxidized silicon (SiO2) substrates is faster than of Au nanoparticles. At high temperature annealing we also find that the aggregation of Pt nanoparticles on the substrate surface is insignificant. As a result, the Pt nanoparticles are uniformly engulfed into the substrates, leading to an opportunity for patterning dense nanopore arrays. Moreover, the use of oxidized Si substrates enables us to precisely control the depth of the nanopores since the engulfment of Pt nanoparticles stops at a short distance above the SiO x /Si interface. After subsequent etching steps, a membrane with dense nanopore through-holes with diameters down to sub-30 nm is obtained. With its simple operation and high controllability, this fabrication method provides an alternative for rapid patterning of dense arrays of solid-state nanopores at low-cost.

Improving the Resolution of 3D-Printed Molds for Microfluidics by Iterative Casting-Shrinkage Cycles.

Breaking through technical barriers and cost reduction are critical issues for the development of microfluidic devices, and both rely greatly on the innovation of fabrication techniques and use of new materials. The application of 3D printing definitely accelerated the prototyping of microfluidic chips by its versatility and functionality. However, the resolution of existing 3D printing techniques is still far below that of lithography, which makes it difficult to work on the scale of single cells and near impossible for single molecule work. In this paper, we present a facile way to increase the resolution of 3D printed microstructures to minimally 4 µm by casting-shrinkage cycles of a polyurethane (PU) polymer. A water-PU liquid mixture poured on a 3D printed template quickly solidifies replicating the structures, which then isometrically shrink to half its size after solvent evaporation, downscaling the replicated structures. By repeating the casting-shrinkage cycles, we could downscale the (sub)millimeter structures of 3D printed structures on demand, until the working limit posed by the polymer properties, which we demonstrate by fabricating a micromixer. Moreover, we can even fabricate microfluidic chips from millimeter-scale manually assembled templates, fully independent of any micromachining facilities, significantly reducing the technical barriers and costs, thus opening up the microfluidics field to low-resource areas.

Integrating Internal Standards into Disposable Capillary Electrophoresis Devices To Improve Quantification.

To improve point-of-care quantification using microchip capillary electrophoresis (MCE), the chip-to-chip variabilities inherent in disposable, single-use devices must be addressed. This work proposes to integrate an internal standard (ISTD) into the microchip by adding it to the background electrolyte (BGE) instead of the sample-thus eliminating the need for additional sample manipulation, microchip redesigns, and/or system expansions required for traditional ISTD usage. Cs and Li ions were added as integrated ISTDs to the BGE, and their effects on the reproducibility of Na quantification were explored. Results were then compared to the conclusions of our previous publication which used Cs and Li as traditional ISTDs. The in-house fabricated microchips, electrophoretic protocols, and solution matrixes were kept constant, allowing the proposed method to be reliably compared to the traditional method. Using the integrated ISTDs, both Cs and Li improved the Na peak area reproducibility approximately 2-fold, to final RSD values of 2.2-4.7% (n = 900). In contrast (to previous work), Cs as a traditional ISTD resulted in final RSDs of 2.5-8.8%, while the traditional Li ISTD performed poorly with RSDs of 6.3-14.2%. These findings suggest integrated ISTDs are a viable method to improve the precision of disposable MCE devices-giving matched or superior results to the traditional method in this study while neither increasing system cost nor complexity.

Ion-Step Method for Surface Potential Sensing of Silicon Nanowires.

This paper presents a novel stimulus-response method for surface potential sensing of silicon nanowire (Si-NW) field-effect transistors. When an "ion-step" from low to high ionic strength is given as a stimulus to the gate oxide surface, an increase of double layer capacitance is therefore expected. Thus, a change of conductance through the Si-NWs is measured. The surface potential on the Si-NW gate is changed from negative for a bare SiO2 surface to neutral/positive when there is poly-l-lysine adsorption at certain pH, which also indicates a shift of point-of-zero charge pH after surface modification. This change is measured by a drop of current variation at the ion-step. The ion-step is performed to the Si-NW through a polydimethylsiloxane microfluidic chip with automatic sample switching. A reduction of the ion-step response from 2 nA to almost zero at pH 5.0 is observed by increasing the potassium ion concentration from 10 mM to 50 mM, which corresponds to a surface potential change of ∼12 mV. We show that this method can be used as an alternative method for surface potential sensing, making it less sensitive to drift.

Nanopore fabrication by heating Au particles on ceramic substrates.

We found that gold nanoparticles, when heated to close to their melting point on substrates of amorphous SiO2 or amorphous Si3N4, move perpendicularly into the substrate. Dependent on applied temperatures, particles can become buried or leave nanopores of extreme aspect ratio (diameter ≅ 25 nm, length up to 800 nm). The process can be understood as driven by gold evaporation and controlled by capillary forces and can be controlled by temperature programming and substrate choice.

Improving chip-to-chip precision in disposable microchip capillary electrophoresis devices with internal standards.

To realize portable systems for routine measurements in point-of-care settings, MCE methods are required to be robust across many single-use chips. While it is well-known internal standards (ISTDs) improve run-to-run precision, a systematic investigation is necessary to determine the significance of chip-to-chip imprecision in MCE and how ISTDs account for it. This paper addresses this question by exploring the reproducibility of Na quantification across six basic, in-house fabricated microchips. A dataset of 900 electrophoerograms was collected from analyzing five concentrations of NaCl with two ISTDs (CsCl and LiCl). While both improved the peak area reproducibility, the Na/Cs ratio was superior to the Na/Li ratio (improving the RSD by a factor of 2-4, depending on the Na concentration). We attribute this to the significant variation in microchannel surface properties, which was accounted for by cesium but not lithium. Microchip dimension and detector variations were only a few percent, and could be improved through commercial fabrication over in-house made microchips. These results demonstrate that ISTDs not only correct for intrachip imprecision, but are also a viable means to correct for chip-to-chip imprecision inherent in disposable, point-of-care MCE devices. However, as expected, the internal standard must be carefully chosen.

Photopatterning of Hydrogel Microarrays in Closed Microchips.

To date, optical lithography has been extensively used for in situ patterning of hydrogel structures in a scale range from hundreds of microns to a few millimeters. The two main limitations which prevent smaller feature sizes of hydrogel structures are (1) the upper glass layer of a microchip maintains a large spacing (typically 525 µm) between the photomask and hydrogel precursor, leading to diffraction of UV light at the edges of mask patterns, (2) diffusion of free radicals and monomers results in irregular polymerization near the illumination interface. In this work, we present a simple approach to enable the use of optical lithography to fabricate hydrogel arrays with a minimum feature size of 4 µm inside closed microchips. To achieve this, we combined two different techniques. First, the upper glass layer of the microchip was thinned by mechanical polishing to reduce the spacing between the photomask and hydrogel precursor, and thereby the diffraction of UV light at the edges of mask patterns. The polishing process reduces the upper layer thickness from ∼525 to ∼100 µm, and the mean surface roughness from 20 to 3 nm. Second, we developed an intermittent illumination technique consisting of short illumination periods followed by relatively longer dark periods, which decrease the diffusion of monomers. Combination of these two methods allows for fabrication of 0.4 × 10(6) sub-10 µm sized hydrogel patterns over large areas (cm(2)) with high reproducibility (∼98.5% patterning success). The patterning method is tested with two different types of photopolymerizing hydrogels: polyacrylamide and polyethylene glycol diacrylate. This method enables in situ fabrication of well-defined hydrogel patterns and presents a simple approach to fabricate 3-D hydrogel matrices for biomolecule separation, biosensing, tissue engineering, and immobilized protein microarray applications.

Nanoscale Chemical Diversity of Coke Deposits on Nanoprinted Metal Catalysts Visualized by Tip-Enhanced Raman Spectroscopy.

Coke formation is the prime cause of catalyst deactivation, where undesired carbon wastes block the catalyst surface and hinder further reaction in a broad gamut of industrial chemical processes. Yet, the origins of coke formation and their distribution across the catalyst remain elusive, obstructing the design of coke-resistant catalysts. Here, the first-time application of tip-enhanced Raman spectroscopy (TERS) is demonstrated as a nanoscale chemical probe to localize and identify coke deposits on a post-mortem metal nanocatalyst. Monitoring coke at the nanoscale circumvents bulk averaging and reveals the local nature of coke with unmatched detail. The nature of coke is chemically diverse and ranges from nanocrystalline graphite to disordered and polymeric coke, even on a single nanoscale location of a top-down nanoprinted SiO2 -supported Pt catalyst. Surprisingly, not all Pt is an equal producer of coke, where clear isolated coke "hotspots" are present non-homogeneously on Pt which generate large amounts of disordered coke. After their formation, coke shifts to the support and undergoes long-range transport on the surrounding SiO2 surface, where it becomes more graphitic. The presented results provide novel guidelines to selectively free-up the coked metal surface at more mild rejuvenation conditions, thus securing the long-term catalyst stability.

An optical aptasensor for real-time quantification of endotoxin: From ensemble to single-molecule resolution.

Endotoxin is a deadly pyrogen, rendering it crucial to monitor with high accuracy and efficiency. However, current endotoxin detection relies on multistep processes that are labor-intensive, time-consuming, and unsustainable. Here, we report an aptamer-based biosensor for the real-time optical detection of endotoxin. The endotoxin sensor exploits the distance-dependent scattering of gold nanoparticles (AuNPs) coupled to a gold nanofilm. This is enabled by the conformational changes of an endotoxin-specific aptamer upon target binding. The sensor can be used in an ensemble mode and single-particle mode under dark-field illumination. In the ensemble mode, the sensor is coupled with a microspectrometer and exhibits high specificity, reliability (i.e., linear concentration to signal profile in logarithmic scale), and reusability for repeated endotoxin measurements. Individual endotoxins can be detected by monitoring the color of single AuNPs via a color camera, achieving single-molecule resolution. This platform can potentially advance endotoxin detection to safeguard medical, food, and pharmaceutical products.

Application of generalized dispersion theory to vortex chromatography.

Acoustically induced secondary flows are applied to enhance lateral mass transfer beyond the relatively slow diffusion. This has the goal to reduce convective axial dispersion and the resulting band broadening which, in turn, limits the performance of column chromatography. Traditional approaches based on Taylor-Aris model are limited to one-dimensional rectilinear (unidirectional) tube- or channel-flows. We therefore apply the generalized dispersion theory (GDT) allowing for prediction of the dependence of potentially improved performance on the characteristics of the induced secondary flow, channel geometry and solute properties as well as providing qualitative physical insight into the role of lateral flows. Results corroborate agreement with our experimental observations (residual standard deviation, Sres = 3.88) and demonstrate the advantage of applying GDT relative to 3D time-dependent simulations.

Electrokinetic DNA transport in 20 nm-high nanoslits: evidence for movement through a wall-adsorbed.

The electrokinetic transport behavior of λ-DNA (48 kbp) in 20 nm-high fused-silica nanoslits in the presence of short-chain PVP is investigated. Mobility and video data show a number of phenomena that are typical of DNA transport through gels or polymer solutions, thus indicative of rigid migration obstacles in the DNA pathway. Calculations show that a several nanometer thin layer of wall-adsorbed PVP ('nano-gel') can provide such a rigid obstacle matrix to the DNA. Such ultrathin wall-adsorbed polymer layers represent a new type of matrix for electrokinetic DNA separation.

Nanofluidics and the chemical potential applied to solvent and solute transport.

The properties and behavior of solvents and solutes inside a nanofluidic structure are not the same as in the bulk solution, but instead are strongly determined by the interactions of solvent and solute molecules with the walls of the structure. These interactions give rise to nanometre-scale boundary layers where the properties can strongly differ from the bulk. The chemical potential provides a convenient tool to describe these boundary layers, and as such is the focus of this tutorial review. The chemical potential of a solution component describes its energy level, which in this boundary layer is strongly influenced by the various interfacial forces between molecules in the wall and in the solution. These forces vary with distance and have a certain limited spatial range, which is reflected in the composition and thickness of the boundary layer in which both solute and solvent concentrations differ from their bulk values. We will consider a variety of solutes such as ions, uncharged molecules and gases, and surfaces that are both hydrophilic and hydrophobic. The boundary layer is oriented normal to the surface, but external forces can also be applied in parallel to the surface. Many interesting different nanofluidic transport phenomena then result, which will also be briefly mentioned in this tutorial review. By this common approach of using the chemical potential for nanofluidic systems of different composition, we aim to bring out the conceptual similarity between the different types of boundary layers and the different transport processes they can give rise to. Finally, as much as possible we will always mention (potential) real-life applications.

A CRISPR/Cas12a-assisted in vitro diagnostic tool for identification and quantification of single CpG methylation sites.

The excellent specificity and selectivity of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/associated nuclease (Cas) is determined by CRISPR RNA's (crRNA's) interchangeable spacer sequence, as well as the position and number of mismatches between target sequence and the crRNA sequence. Some diseases are characterized by epigenetic alterations rather than nucleotide changes, and are therefore unsuitable for CRISPR-assisted sensing methods. Here we demonstrate an in vitro diagnostic tool to discriminate single CpG site methylation in DNA by the use of methylation-sensitive restriction enzymes (MSREs) followed by Cas12a-assisted sensing. Non-methylated sequences are digested by MSREs, resulting in fragmentation of the target sequence that influences the R-loop formation between crRNA and target DNA. We show that fragment size, fragmentation position and number of fragments influence the subsequent collateral trans-cleavage activity towards single stranded DNA (ssDNA), enabling deducting the methylation position from the cleavage activity. Utilizing MSREs in combination with Cas12a, single CpG site methylation levels of a cancer gene are determined. The modularity of both Cas12a and MSREs provides a high level of versatility to the Cas12a-MSRE combined sensing method, which opens the possibility to easily and rapidly study single CpG methylation sites for disease detection.