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.

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.

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.

Inducing AC-electroosmotic flow using electric field manipulation with insulators.

Classically, the configuration of electrodes (conductors) is used as a means to determine AC-electroosmotic flow patterns. In this paper, we use the configuration of insulator materials to achieve AC-electroosmotic flow patterning in a novel approach. We apply AC electric fields between parallel electrodes situated on the top and bottom of a microfluidic channel and separated by an insulating material. Channels of various cross-sectional shapes (e.g. rectangular and parallelogram) were fabricated by shaping the insulating material between the electrodes. We found that vortex flow patterns are induced depending on the cross-sectional shape of the channel. A bell-shaped design with non-orthogonal corners gave rise to 2 vortices, whereas in a channel with a parallelogram shaped cross-section, only a single vortex was observed. The vortices were experimentally observed by analysing the 3D trajectories of fluorescent microparticles. From a theoretical analysis, we conclude that flow shaping is primarily caused by shaping the electrical field lines in the channel.

Bottom-Up Assembled Photonic Crystals for Structure-Enabled Label-Free Sensing.

Photonic crystals (PhCs) display photonic stop bands (PSBs) and at the edges of these PSBs transport light with reduced velocity, enabling the PhCs to confine and manipulate incident light with enhanced light-matter interaction. Intense research has been devoted to leveraging the optical properties of PhCs for the development of optical sensors for bioassays, diagnosis, and environmental monitoring. These applications have furthermore benefited from the inherently large surface area of PhCs, giving rise to high analyte adsorption and the wide range of options for structural variations of the PhCs leading to enhanced light-matter interaction. Here, we focus on bottom-up assembled PhCs and review the significant advances that have been made in their use as label-free sensors. We describe their potential for point-of-care devices and in the review include their structural design, constituent materials, fabrication strategy, and sensing working principles. We thereby classify them according to five sensing principles: sensing of refractive index variations, sensing by lattice spacing variations, enhanced fluorescence spectroscopy, surface-enhanced Raman spectroscopy, and configuration transitions.

Patterning Biological Gels for 3D Cell Culture inside Microfluidic Devices by Local Surface Modification through Laminar Flow Patterning.

Microfluidic devices are used extensively in the development of new in vitro cell culture models like organs-on-chips. A typical feature of such devices is the patterning of biological hydrogels to offer cultured cells and tissues a controlled three-dimensional microenvironment. A key challenge of hydrogel patterning is ensuring geometrical confinement of the gel, which is generally solved by inclusion of micropillars or phaseguides in the channels. Both of these methods often require costly cleanroom fabrication, which needs to be repeated even when only small changes need be made to the gel geometry, and inadvertently expose cultured cells to non-physiological and mechanically stiff structures. Here, we present a technique for facile patterning of hydrogel geometries in microfluidic chips, but without the need for any confining geometry built into the channel. Core to the technique is the use of laminar flow patterning to create a hydrophilic path through an otherwise hydrophobic microfluidic channel. When a liquid hydrogel is injected into the hydrophilic region, it is confined to this path by the surrounding hydrophobic regions. The various surface patterns that are enabled by laminar flow patterning can thereby be rendered into three-dimensional hydrogel structures. We demonstrate that the technique can be used in many different channel geometries while still giving the user control of key geometric parameters of the final hydrogel. Moreover, we show that human umbilical vein endothelial cells can be cultured for multiple days inside the devices with the patterned hydrogels and that they can be stimulated to migrate into the gel under the influence of trans-gel flows. Finally, we demonstrate that the patterned gels can withstand trans-gel flow velocities in excess of physiological interstitial flow velocities without rupturing or detaching. This novel hydrogel-patterning technique addresses fundamental challenges of existing methods for hydrogel patterning inside microfluidic chips, and can therefore be applied to improve design time and the physiological realism of microfluidic cell culture assays and organs-on-chips.

Reduction of Taylor-Aris dispersion by lateral mixing for chromatographic applications.

Chromatographic columns are suffering from Taylor-Aris dispersion, especially for slowly diffusing molecules such as proteins. Since downscaling the channel size to reduce Taylor-Aris dispersion meets fundamental pressure limitations, new strategies are needed to further improve chromatography beyond its current limits. In this work we demonstrate a method to reduce Taylor-Aris dispersion by lateral mixing in a newly designed silicon AC-electroosmotic flow mixer. We obtained a reduction in κaris by a factor of three in a 40 µm × 20 µm microchannel, corresponding to a plate height gain of 2 to 3 under unretained conditions at low to high Pe values. We also demonstrate an improvement of a reverse-phase chromatographic separation of coumarins.

Point-of-care CRISPR/Cas nucleic acid detection: Recent advances, challenges and opportunities.

With the trend of moving molecular tests from clinical laboratories to on-site testing, there is a need for nucleic acid based diagnostic tools combining the sensitivity, specificity and flexibility of established diagnostics with the ease, cost effectiveness and speed of isothermal amplification and detection methods. A promising new nucleic acid detection method is Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated nuclease (Cas)-based sensing. In this method Cas effector proteins are used as highly specific sequence recognition elements that can be combined with many different read-out methods for on-site point-of-care testing. This review covers the technical aspects of integrating CRISPR/Cas technology in miniaturized sensors for analysis on-site. We start with a short introduction to CRISPR/Cas systems and the different effector proteins and continue with reviewing the recent developments of integrating CRISPR sensing in miniaturized sensors for point-of-care applications. Finally, we discuss the challenges of point-of-care CRISPR sensing and describe future research perspectives.

Plasmonic Nanocrystal Arrays on Photonic Crystals with Tailored Optical Resonances.

Hierarchical plasmonic-photonic microspheres (PPMs) with high controllability in their structures and optical properties have been explored toward surface-enhanced Raman spectroscopy. The PPMs consist of gold nanocrystal (AuNC) arrays (3rd-tier) anchored on a hexagonal nanopattern (2nd-tier) assembled from silica nanoparticles (SiO2NPs) where the uniform microsphere backbone is termed the 1st-tier. The PPMs sustain both photonic stop band (PSB) properties, resulting from periodic SiO2NP arrangements of the 2nd-tier, and a surface plasmon resonance (SPR), resulting from AuNC arrays of the 3rd-tier. Thanks to the synergistic effects of the photonic crystal (PC) structure and the AuNC array, the electromagnetic (EM) field in such a multiscale composite structure can tremendously be enhanced at certain wavelengths. These effects are demonstrated by experimentally evaluating the Raman enhancement of benzenethiol (BT) as a probe molecule and are confirmed via numerical simulations. We achieve a maximum SERS enhancement factor of up to ∼108 when the resonances are tailored to coincide with the excitation wavelength by suitable structural modifications.

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.

Droplet encapsulation of electrokinetically-focused analytes without loss of resolution.

Lab-on-chip electrokinetic focusing and separation techniques are widely used in several scientific fields. In a number of cases, these techniques have been combined with a selective analyte extraction for off-chip analysis. Nevertheless, the usability of the extracts is limited by diffusion which reduces the separation resolution. In this paper we propose the integration of a droplet generator capable of continuous or on-demand generation and extraction of electrokinetically separated and focused analytes. We demonstrate the selective droplet extraction of model analytes separated and concentrated via ion concentration polarization focusing (ICPF). We report extracted droplets with 1000-fold increased concentration. Importantly, the droplet generator does not interrupt the ICPF process making it suitable for integration with the majority of electrokinetic separation techniques.

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.

Dimension-reconfigurable bubble film nanochannel for wetting based sensing.

Dimensions and surface properties are the predominant factors for the applications of nanofluidic devices. Here we use a thin liquid film as a nanochannel by inserting a gas bubble in a glass capillary, a technique we name bubble-based film nanofluidics. The height of the film nanochannel can be regulated by the Debye length and wettability, while the length independently changed by applied pressure. The film nanochannel behaves functionally identically to classical solid state nanochannels, as ion concentration polarizations. Furthermore, the film nanochannels can be used for label-free immunosensing, by principle of wettability change at the solid interface. The optimal sensitivity for the biotin-streptavidin reaction is two orders of magnitude higher than for the solid state nanochannel, suitable for a full range of electrolyte concentrations. We believe that the film nanochannel represents a class of nanofluidic devices that is of interest for fundamental studies and also can be widely applied, due to its reconfigurable dimensions, low cost, ease of fabrication and multiphase interfaces.

Organ-on-a-chip: the next generation platform for risk assessment of radiobiology.

Organ-on-a-chip devices have been widely used in biomedical science and technology, for example for experimental regenerative medicine and precision healthcare. The main advantage of organ-on-a-chip technology is the facility to build a specific human model that has functional responses on the level of organs or tissues, thereby avoiding the use of animal models, as well as greatly improving new drug discovery processes for personal healthcare. An emerging application domain for organs-on-chips is the study of internal irradiation for humans, which faces the challenges of the lack of a clear model for risk estimation of internal irradiation. We believe that radiobiology studies will benefit from organ-on-a-chip technology by building specific human organ/tissues in vitro. In this paper, we briefly reviewed the state-of-the-art in organ-on-a-chip research in different domains, and conclude with the challenges of radiobiology studies at internal low-dose irradiation. Organ-on-a-chip technology has the potential to significantly improve the radiobiology study as it can mimic the function of human organs or tissues, and here we summarize its potential benefits and possible breakthrough areas, as well as its limitations in internal low-dose radiation studies.

Plasmonic Bubble Nucleation and Growth in Water: Effect of Dissolved Air.

Under continuous laser irradiation, noble metal nanoparticles immersed in water can quickly heat up, leading to the nucleation of so-called plasmonic bubbles. In this work, we want to further understand the bubble nucleation and growth mechanism. In particular, we quantitatively study the effect of the amount of dissolved air on the bubble nucleation and growth dynamics, both for the initial giant bubble, which forms shortly after switching on the laser and is mainly composed of vapor, and for the final life phase of the bubble, during which it mainly contains air expelled from water. We found that the bubble nucleation temperature depends on the gas concentration: the higher the gas concentration, the lower the bubble nucleation temperature. Also, the long-term diffusion-dominated bubble growth is governed by the gas concentration. The radius of the bubbles grows as R(t) ∝ t 1/3 for air-equilibrated and air-oversaturated water. In contrast, in partially degassed water, the growth is much slower since, even for the highest temperature we achieve, the water remains undersaturated.

Continuous focusing, fractionation and extraction of anionic analytes in a microfluidic chip.

Electrokinetic focusing and separation methods, specifically ion concentration polarization focusing (ICPF), provide a very powerful and easy to use analytical tool for several scientific fields. Nevertheless, the concentrated and separated analytes are effectively trapped inside the chip in picoliter volumes. In this article we propose an ICPF device that allows continuous and selective extraction of the focused analytes. A theoretical background is presented to understand the dynamics of the system and a 1D model was developed that describes the general behavior of the system. We demonstrate the selective extraction of three fluorescent model anionic analytes and we report selective extraction of the analytes at a 300-fold increased concentration.

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.

Wafer-scale fabrication of high-quality tunable gold nanogap arrays for surface-enhanced Raman scattering.

We report a robust and high-yield fabrication method for wafer-scale patterning of high-quality arrays of dense gold nanogaps, combining displacement Talbot lithography based shrink-etching with dry etching, wet etching, and thin film deposition techniques. By using the self-sharpening of <111>-oriented silicon crystal planes during the wet etching process, silicon structures with extremely smooth nanogaps are obtained. Subsequent conformal deposition of a silicon nitride layer and a gold layer results in dense arrays of narrow gold nanogaps. Using this method, we successfully fabricate high-quality Au nanogaps down to 10 nm over full wafer areas. Moreover, the gap spacing can be tuned by changing the thickness of deposited Au layers. Since the roughness of the template is minimized by the crystallographic etching of silicon, the roughness of the gold nanogaps depends almost exclusively on the roughness of the sputtered gold layers. Additionally, our fabricated Au nanogaps show a significant enhancement of surface-enhanced Raman scattering (SERS) signals of benzenethiol molecules chemisorbed on the structure surface, at an average enhancement factor up to 1.5 × 106.