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openaire: EC/H2020/725513/EU//SuperRepel A myriad of natural surfaces such as plant leaves and insect wings can repel water and remain unwetted inspiring scientists and engineers to develop water-repellent surfaces for various applications. Those natural and artificial water-repellent surfaces are typically opaque, containing micro- and nano-roughness, and their wetting properties are determined by the details at the actual liquid-solid interface. However, a generally applicable way to directly observe moving contact lines on opaque water-repellent surfaces is missing. Here, we show that the advancing and receding contact lines and corresponding contact area on micro- and nano-rough water-repellent surfaces can be readily and reproducibly quantified using a transparent droplet probe. Combined with a conventional optical microscope, we quantify the progression of the apparent contact area and apparent contact line irregularity in different types of superhydrophobic silicon nanograss surfaces. Contact angles near 180° can be determined with an uncertainty as low as 0.2°, that a conventional contact angle goniometer cannot distinguish. We also identify the pinning/depinning sequences of a pillared model surface with excellent repeatability and quantify the progression of the apparent contact interface and contact angle of natural plant leaves with irregular surface topography.

Brain-on-Chip devices, which enable on-chip cultures of neurons to simulate brain functions, are receiving tremendous attention to meet the demands of both fundamental research and clinical trials. Thus, miniaturized sensors are being developed to accomplish real-time monitoring of neurotransmitters, which are the signaling molecules between neurons and benchmarks for neuron network operation. Electrochemical sensors are promising candidates for detecting a critical neurotransmitter, dopamine (DA), a biomarker for Parkinson’s disease. A key advantage of electrochemical sensors is their capability of fast, real-time recording. However, current state-of-the-art electrochemical DA sensors are suffering from various issues like limited sensitivity and cumbersome fabrication. Here, we report a novel route in monolithically microfabricating vertically aligned carbon nanofiber (VACNF) electrochemical DA microsensors by an anti-blistering slow cooling process. Thanks to the new microfabrication process, we are able to achieve VACNF microsensors with complete insulation and large surface areas. The champion device shows extremely high sensitivity of 4.52×104 μA/μM · cm2, which is at least two-orders-of-magnitude higher than current devices, and a highly competitive limit of detection of 0.243 nM. These figures of merit will open new windows for future applications of Brain-on-Chip devices such as electrochemical recording from a single neuron.

A.P. and C.F. gratefully acknowledge the Finnish Cultural Foundation and the Emil Aaltonen Foundation for financial support. The work utilized the facilities of the Otanano and RAMI national research infrastructure and the cleanroom facilities of Micronova. J.V. acknowledges with gratitude funding from the Academy of Finland (Decision Number: 322214) and Flagship Programme, Photonics Research and Innovation (PREIN, Decision Number: 320167). Herein, a simple hierarchical surface patterning method is presented by effectively combining buckling instability and azopolymer-based surface relief grating inscription. In this technique, submicron patterns are achieved using azopolymers, whereas the microscale patterns are fabricated by subsequent thermal shrinkage. The wetting characterization of various topographically patterned surfaces confirms that the method permits tuning of contact angles and choosing between isotropic and anisotropic wetting. Altogether, this method allows efficient fabrication of hierarchical surfaces over several length scales in relatively large areas, overcoming some limitations of fabricating multiscale roughness in lithography and also methods of creating merely random patterns, such as black silicon processing or wet etching of metals. The demonstrated fine-tuning of the surface patterns may be useful in optimizing surface-related material properties, such as wetting and adhesion, producing substrates that are of potentialinterest in mechanobiology and tissue engineering.

Tissue oxygen levels are known to be critical to regulation of many cellular processes, including the hepatic metabolism of therapeutic drugs, but its impact is often ignored in in vitro assays. In this study, the material-induced oxygen scavenging property of off-stoichiometric thiol-enes (OSTE) was exploited to create physiologically relevant oxygen concentrations in microfluidic immobilized enzyme reactors (IMERs) incorporating human liver microsomes. This could facilitate rapid screening of, for instance, toxic drug metabolites possibly produced in hypoxic conditions typical for many liver injuries. The mechanism of OSTE-induced oxygen scavenging was examined in depth to enable precise adjustment of the on-chip oxygen concentration with the help of microfluidic flow. The oxygen scavenging rate of OSTE was shown to depend on the type and the amount of the thiol monomer used in the bulk composition, and the surface-to-volume ratio of the chip design, but not on the physical or mechanical properties of the bulk. Our data suggest that oxygen scavenging takes place at the polymer-liquid interface, likely via oxidative reactions of the excess thiol monomers released from the bulk with molecular oxygen. Based on the kinetic constants governing the oxygen scavenging rate in OSTE microchannels, a microfluidic device comprising monolithically integrated oxygen depletion and IMER units was designed and its performance validated with the help of oxygen-dependent metabolism of an antiretroviral drug, zidovudine, which yields a cytotoxic metabolite under hypoxic conditions.

International audience; Different types of carbon materials are biocompatible with neural cells and can promote maturation. The mechanism of this effect is not clear. Here we have tested the capacity of a carbon material composed of amorphous sp 3 carbon backbone, embedded with a percolating network of sp 2 carbon domains to sustain neuronal cultures. We found that cortical neurons survive and develop faster on this novel carbon material. After 3 days in culture, there is a precocious increase in the frequency of neuronal activity and in the expression of maturation marker KCC2 on carbon films as compared to a commonly used glass surface. Accelerated development is accompanied by a dramatic increase in neuronal dendrite arborization. The mechanism for the precocious maturation involves the activation of intracellular calcium oscillations by the carbon material already after 1 day in culture. Carbon-induced oscillations are independent of network activity and reflect intrinsic spontaneous activation of developing neurons. Thus, these results reveal a novel mechanism for carbon material-induced neuronal survival and maturation. Brain trauma as well as neurodegenerative diseases are the leading cause of irreversible disability and low quality of life in the elderly population 1. A way to combat neurodegeneration is to promote reparation of neuronal networks, rewiring of neuronal connections, and eventual restoration or substitution of the lost functionality 2. One putative therapeutic avenue is providing scaffolds-special materials that support targeted differentiation of neuronal stem cells and neurite outgrowth of regenerating neurons. Carbon-derived materials possess numerous properties that make them usable as scaffolds 3. Carbon nanotubes (CNTs) and graphene are among the most studied carbon materials for biological applications. These materials enhance neuronal stem cell differentiation 4-6 , as well as promote neuronal survival, neuronal activity, and neuronal process outgrowth 7-13. Neurons cultured on CNTs have increased levels of neuronal K +-Cl − cotransporter KCC2, a key component in the functional maturation of inhibitory synaptic 14 and glutamatergic 15-17 transmission. Downregulation of this protein is also implicated in reactive plasticity following brain trauma 18. CNTs have been suggested to improve the electrical responsiveness of neurons by facilitating local electronic shortcuts between somas and dendrites 19. The ability of CNTs to form tight contacts with neurons is beneficial for neuron-electrode interfaces 20-24. 3D gra-phene substrates support growth and differentiation of neurons 25-27 that in combination with anti-inflammatory properties 28,29 makes graphene a next-generation neuronal tissue scaffold. Despite the importance of novel carbon materials for future engineering, we do not fully understand the mechanisms underlining the trophic action of carbon scaffolds. In this work, we propose a novel mechanism by which a new type of sputtered carbon material accelerates neuronal maturation. The carbon film material consists of conducting nanoscale sp 2 carbon islands embedded in a diamond-like sp 3 carbon matrix. We demonstrate OPEN

Superomniphobic surfaces that repel liquids of extremely low surface tension rely on carefully fabricated doubly re-entrant topographies, typically made by silicon deep reactive ion etching technology. However, previously published processes have depended on critically timed etching steps, which are difficult to downscale. We present a scalable process that eliminates the critically timed etching steps. It is based on the use of silicon-on-insulator wafers and a silicon oxide foot of the micropillar, which makes the isotropic silicon release step non-critical. The process allows easy downscaling of pillars from 20 $\mu \text{m}$ to 10 $\mu \text{m}$ and $5~\mu \text{m}$ . The downscaling increases the stability of the Cassie state. Based on the process, we are able to create superomniphobic surfaces that sustain perfluorohexane (FC-72), which has the lowest surface tension of the known liquids at room temperature ( $\gamma _{\text {lv}}=11.91$ mN/m at 20 °C), in the Cassie state at droplet diameters down to 200 micrometers. These are the smallest perfluorohexane droplets repelled to date. [2019-0207]

openaire: EC/H2020/824070/EU//CONNECT Funding Information: The authors thank Dr. Ville Rontu for valuable discussions in ALD process, Niklas Wester and Antti Peltonen for help with preparing electrode samples. The work utilized the fabrication and characterization facilities of Aalto University, cleanroom facilities of Micronova micro and nanofabrication center, and Nanotalo-Nanomicroscopy Center (Aalto-NMC). This project has received funding from the Horizon 2020 Framework Programme Project : 824070 — CONNECT. Publisher Copyright: © 2021 Titanium nitride (TiN) is widely utilized in microelectrode array fabrication for electrophysiological recordings due to its relatively low noise levels, long term stability, and exceptional biocompatibility. Atomic layer deposition (ALD) is a well-established approach for the fabrication of TiN thin films, offering great control over the thickness and properties of the films. Although, advanced procedures have been reported to develop micro and nanostructured electrodes, TiN thin film has yet not been widely applied as electrode material for electrochemical sensing, and characterization of these ALD fabricated TiN films is lacking. Here, we study the use of TiN thin films as electrochemical signal transducer for neurotransmitter dopamine (DA) detection with emphasis on investigation the effect of oxygen functionalities on the electrochemical performance of the films. We find that in order to have high enough sensitivity and selectivity the electrode material used to realize the measurements must be modified.In this work, we described TiN thin film surface modification through hybridization of microfabrication and nanocomposite approaches. Because of its good adhesion, TiN is considered as a high interesting support for Pt catalyst. Therefore, ALD was used to deposit TiN thin films and to design Pt nanoparticles (Pt NPs) on highly conductive boron-doped silicon. Further modification was done through multiwalled carbon nanotubes (MWCNTs), which were immobilized on Pt NPs/ TiN hybrid electrode by Nafion film. Synergistic effect of Pt NPs, MWCNTs, and Nafion film caused significant increase in the electrocatalytic activity towards DA electrooxidation. The proposed sensor material was characterized by focused ion beam milling combined with scanning electron microscopy (FIB-SEM), X-ray photoelectron spectroscopy (XPS), transmission electron microscopy (TEM), spectroscopic ellipsometry, contact angle measurement and its electrochemical behavior was studied by cyclic voltammetry (CV). CV technique was applied for sensing DA in the range of 0.05–35 µM (R2 = 0.990) with a good sensitivity (16.31 μA/μM) and a low detection limit of 40 nM (for S/N = 3). The developed sensor exhibited reasonable response time and good stability.

Funding Information: This work was supported by the AaltoENG doctoral programme of the School of Engineering of the Aalto University. Publisher Copyright: © 2021 The Author(s) Copyright: Copyright 2021 Elsevier B.V., All rights reserved. This study experimentally examines the relationship between fibre surface roughness, its wettability, and the microstructure of the cement paste close to the fibre. Results indicate that the amount of water that adheres to or is retained by the fibre surface grows with the increase in roughness. The distributions of porosity and of unhydrated cement grains, which were produced through analysis of BSE images with a k-means clustering algorithm up to 100μm from fibre surface, revealed a decrease in both large pores and porosity when roughness increased but not a clear effect on the distribution of unhydrated cement grains.

Flexible and conductive silver electrodes were fabricated by inkjet printing on several different compositions of thiol-ene polymers. Conductive electrodes with resistivity down to 30 µΩcm and good adhesion of the electrodes were obtained by optimizing the printing parameters. The maximum printing resolution was 100 µm lines and 80 µm gaps between the lines. Printing on top of cross-linked off-stoichiometric thiol-ene polymer was tested for compositions ranging from 30 % thiol excess to 5 % allyl (‘ene’) excess. The roughness off the thiol-ene surfaces was shown to greatly improve the quality of the printed electodes: consistently high yield of conductive electrodes was obtained on rough surfaces (roughness ≈ 1 µm), whereas on smooth surfaces the electrodes were often cracked. The lowest resistivity values were obtained on electrodes printed on near stoichiometric thiol-ene substrates. The conductivity of the electrodes was retained after 5 % linear strain and after repeated bending with 1 mm radius of curvature, showing the potential for flexible sensors. The electrodes were also applied to electrical impedance-based monitoring of cell growth on thiol-ene surfaces, which showcased that the electrodes survive stressed cell culture conditions for at least 36 h.

Single cell trapping is demonstrated on superhydrophobic-superhydrophilic patterned microarrays. Superhydrophilic spots that are twice the size of the cells to be trapped are found to be optimal for obtaining single cell trapping. The single cell trapping is based on size exclusivity instead of heavy dilution of the cell suspension and relying on Poisson statistics. The superhydrophobic areas of the array are found to be very resistant toward unwanted adhesion of the cells and thus cleaning steps are not needed after deposition. Based on these properties, 20 mu m superhydrophilic spots are utilized for trapping two types of immune cells, primary peripheral blood mononuclear cells (PBMCs) and THP-1 cells. The cells are trapped from a 10 mu l cell suspension droplet dragged across the surface with different velocities. Single cell trapping efficiencies in the range of 10-30% are shown based on the cell type, the spot type, the seeding velocity and the cell suspension concentration.

openaire: EC/H2020/| 311705/EU//CUMTAS We introduce rapid replica molding of ordered, high-aspect-ratio, thiol-ene micropillar arrays for implementation of microfluidic immobilized enzyme reactors (IMERs). By exploiting the abundance of free surface thiols of off-stoichiometric thiol-ene compositions, we were able to functionalize the native thiol-ene micropillars with gold nanoparticles (GNPs) and these with proteolytic α-chymotrypsin (CHT) via thiol-gold interaction. The micropillar arrays were replicated via PDMS soft lithography, which facilitated thiol-ene curing without the photoinitiators, and thus straightforward bonding and good control over the surface chemistry (number of free surface thiols). The specificity of thiol-gold interaction was demonstrated over allyl-rich thiol-ene surfaces and the robustness of the CHT-IMERs at different flow rates and reaction temperatures using bradykinin hydrolysis as the model reaction. The product conversion rate was shown to increase as a function of decreasing flow rate (increasing residence time) and upon heating of the IMER to physiological temperature. Owing to the effective enzyme immobilization onto the micropillar array by GNPs, no further purification of the reaction solution was required prior to mass spectrometric detection of the bradykinin hydrolysis products and no clogging problems, commonly associated with conventional capillary packings, were observed. The activity of the IMER remained stable for at least 1.5 h (continuous use), suggesting that the developed protocol may provide a robust, new approach to implementation of IMER technology for proteomics research. [Figure not available: see fulltext.].

Additive manufacturing (3D printing) is a disruptive technology that is changing production systems globally. In addition, microfluidic devices are increasingly being used for chemical analysis and continuous production of chemicals. Printing of materials such as polymers and metals is already a reality, but additive manufacturing of glass for microfluidic systems has received minor attention. We characterize microfluidic devices (channel cross-section dimensions down to a scale of 100 mm) that have been produced by additive manufacturing of molten soda-lime glass in tens of minutes and report their mass spectrometric and Raman spectroscopic analysis examples. The functionality of a microfluidic glass microreactor is shown with online mass spectrometric analysis of linezolid synthesis. Additionally, the performance of a direct infusion device is demonstrated by mass spectrometric analysis of drugs. Finally, the excellent optical quality of the glass structures is demonstrated with in-line Raman spectroscopic measurements. Our results promise a bright future for additively manufactured glass microdevices in diverse fields of science.

Stem cell renewal and differentiation are regulated by interactions with the niche. Although multiple cell populations have been identified in distinct anatomical compartments, little is known about niche-specific molecular factors. Using skin as a model system and combining single-cell RNA-seq data analysis, immunofluorescence, and transgenic mouse models, we show that the transmembrane protein embigin is specifically expressed in the sebaceous gland and that the number of embigin-expressing cells is negatively regulated by Wnt. The loss of embigin promotes exit from the progenitor compartment and progression toward differentiation, and also compromises lipid metabolism. Embigin modulates sebaceous niche architecture by affecting extracellular matrix organization and basolateral targeting of monocarboxylate transport. We discover through ligand screening that embigin is a direct fibronectin receptor, binding to the N-terminal fibronectin domain without impairing integrin function. Our results solve the long-standing question of how embigin regulates cell adhesion and demonstrate a mechanism that couples adhesion and metabolism.

The performance of fibre reinforced cementitious composites depends on the interaction between the cementitious matrix and the fibres on the micro-scale. This interaction is affected by the material and surface properties of the fibre and the micro-scale characteristics of the cementitious matrix close to the fibre surface, creating the interfacial transition zone (ITZ). However, the discussions regarding the effect of the surface properties of fibres on the formation of the ITZ and, as a result, the bond strength between the matrix and the fibres are limited. The present paper reports the progress of the ongoing research project, which concentrates on investigating the contact created by the steel fibres with different characteristics of the surface roughness and cementitious matrix. The results of this study can support the development of efficient fibre reinforced cementitious composites.

The performance of fibre reinforced cementitious composites depends on the interaction between the cementitious matrix and the fibres on the micro-scale. This interaction is affected by the material and surface properties of the fibre and the micro-scale characteristics of the cementitious matrix close to the fibre surface, creating the interfacial transition zone (ITZ). However, the discussions regarding the effect of the fibre surface on the formation of the ITZ and, as a result, the bond strength between the matrix and the fibres are limited. The present paper reports the progress of the ongoing research project, which concentrates on investigating the interface created by the steel fibres with different characteristics of the surface roughness and cementitious matrix. The results of this study can support the development of efficient fibre reinforced cementitious composites.

A hypoxic (low oxygen level) microenvironment and nitric oxide paracrine signaling play important roles in the control of both biological and pathological cell responses. In this study, we present a microfluidic chip architecture for nitric oxide delivery under a hypoxic microenvironment in human embryonic kidney cells (HEK-293). The chip utilizes two separate, but interdigitated microfluidic channels. The hypoxic microenvironment was created by sodium sulfite as the oxygen scavenger in one of the channels. The nitric oxide microenvironment was created by sodium nitroprusside as the light-activated nitric oxide donor in the other channel. The solutions are separated from the cell culture by a 30 &micro, m thick gas-permeable, but liquid-impermeable polydimethylsiloxane membrane. We show that the architecture is preliminarily feasible to define the gaseous microenvironment of a cell culture in the 100 &micro, m and 1 mm length scales.

Biofouling imposes a significant threat for sensing probes used in vivo. Antifouling strategies commonly utilize a protective layer on top of the electrode but this may compromise performance of the electrode. Here, we investigated the effect of surface topography and chemistry on fouling without additional protective layers. We have utilized two different carbon materials; tetrahedral amorphous carbon (ta-C) and SU-8 based pyrolytic carbon (PyC) in their typical smooth thin film structure as well as with a nanopillar topography templated from black silicon. The near edge X-ray absorption fine structure (NEXAFS) spectrum revealed striking differences in chemical functionalities of the surfaces. PyC contained equal amounts of ketone, hydroxyl and ether/epoxide groups, while ta-C contained significant amounts of carbonyl groups. Overall, oxygen functionalities were significantly increased on nanograss surfaces compared to the flat counterparts. Neither bovine serum albumin (BSA) or fetal bovine serum (FBS) fouling caused major effects on electron transfer kinetics of outer sphere redox (OSR) probe Ru(NH3)63+ on any of the materials. In contrast, negatively charged OSR probe IrCl62- kinetics were clearly affected by fouling, possibly due to the electrostatic repulsion between redox species and the anionically-charged proteins adsorbed on the electrode and/or stronger interaction of the proteins and positively charged surface. The OSR probe kinetics were less affected by fouling on PyC, probably due to conformational changes of proteins on the surface. Dopamine (DA) was tested as an inner sphere redox (ISR) probe and as expected, the kinetics were heavily dependent on the material; PyC had very fast electron transfer kinetics, while ta-C had sluggish kinetics. DA electron transfer kinetics were heavily affected on all surfaces by fouling (ΔEp increase 30-451%). The effect was stronger on PyC, possibly due to the more strongly adhered protein layer limiting the access of the probe to the inner sphere.

Two‐dimensional (2D) cell cultures have been the primary screening tools to predict drug impacts in vitro for decades. However, owing to the lack of tissue‐specific architecture of 2D cultures, secondary screening using three‐dimensional (3D) cell culture models is often necessary. A microfluidic approach that facilitates side‐by‐side 2D and 3D cell culturing in a single microchannel and thus combines the benefits of both set‐ups in drug screening; that is, the uniform spatiotemporal distributions of oxygen, nutrients, and metabolic wastes in 2D, and the tissue‐like architecture, cell–cell, and cell–extracellular matrix interactions only achieved in 3D. The microfluidic platform is made from an organically modified ceramic material, which is inherently biocompatible and supports cell adhesion (2D culture) and metal adhesion (for integration of impedance electrodes to monitor cell proliferation). To induce 3D spheroid formation on another area, a single‐step lithography process is used to fabricate concave microwells, which are made cell‐repellant by nanofunctionalization (i.e., plasma porosification and hydrophobic coating). Thanks to the concave shape of the microwells, the spheroids produced on‐chip can also be released, with the help of microfluidic flow, for further off‐chip characterization after culturing. In this study, the methodology is evaluated for drug cytotoxicity assessment on human hepatocytes.

We present a simple and facile method for long-term preservation of hydrophilicity of oxygen plasma-hydrophilized poly (dimethylsiloxane) (PDMS) by cold storage. We show that storage under temperature of − 80 °C can maintain superhydrophilicity of plasma-exposed PDMS for at least 100 days. Storage at − 15 °C and at 22 °C room temperature (RT) is shown to exhibit, respectively, about half and full recovery of the original hydrophobicity after 100 days in storage. Furthermore, we investigated the implications of the cold storage for microfluidic applications, the capillary filling rate and the ability of the flow to bypass geometrical obstacles in a microfluidic channel. It is shown that the preservation of capillary filling properties of microchannels is in close agreement with the contact angle (CA) measurements and that the colder the storage temperature, the better the capillary filling capability of the channels is preserved. We ascribe the significantly reduced recovery rate to reduced thermally activated relaxation phenomena such as diminished diffusion of low molecular weight species (LMW) in the polymer matrix at colder temperatures. This is supported by ATR-FTIR measurements of the OH vibration band over time for samples stored at different temperatures.

Carbon-based materials have attracted much attention in biological applications like interfacing electrodes with neurons and cell growth platforms due to their natural biocompatibility and tailorable material properties. Here we have fabricated sputtered carbon thin film electrodes for bioelectrical measurements. Reactive ion etching (RIE) recipes were optimized with Taguchi method to etch the close field unbalanced magnetron sputtered carbon thin film (nanocarbon, nC) consisting of nanoscale crystalline sp(2)-domains in amorphous sp(3)-bonded backbone. Plasma etching processes used gas mixtures of Ar/O-2/SF6/CHF3 for RIE and O-2/SF6 for ICP-RIE. The highest achieved etch rate for nanocarbon was >> 389 nm/min and best chromium etch mask selectivity was 135:1. Biocompatibility of the material was tested with rat neuronal cultures. Next, we fabricated multielectrode arrays (MEA) with carbon recording electrodes and metal wiring. Organotypic brain slices grown on the MEAs were viable and showed characteristic spontaneous electrical network activity. The results demonstrate that interactions with nanocarbon substrate support neuronal survival and maturation of functional neuronal networks. Thus the material can have wide applications in biomedical research.

This work describes aqueous and non-aqueous capillary electrophoresis on thiol-ene-based microfluidic separation devices that feature fully integrated and sharp electrospray ionization (ESI) emitters. The chip fabrication is based on simple and low-cost replica-molding of thiol-ene polymers under standard laboratory conditions. The mechanical rigidity and the stability of the materials against organic solvents, acids and bases could be tuned by adjusting the respective stoichiometric ratio of the thiol and allyl (“ene”) monomers, which allowed us to carry out electrophoresis separation in both aqueous and non-aqueous (methanol- and ethanol-based) background electrolytes. The stability of the ESI signal was generally ≤10% RSD for all emitters. The respective migration time repeatabilities in aqueous and non-aqueous background electrolytes were below 3 and 14% RSD (n = 4-6, with internal standard). The analytical performance of the developed thiol-ene microdevices was shown in mass spectrometry (MS) based analysis of peptides, proteins, and small molecules. The theoretical plate numbers were the highest (1.2–2.4 × 104 m−1) in ethanol-based background electrolytes. The ionization efficiency also increased under non-aqueous conditions compared to aqueous background electrolytes. The results show that replica-molding of thiol-enes is a feasible approach for producing ESI microdevices that perform in a stable manner in both aqueous and non-aqueous electrophoresis.

Here we investigated the electrochemical properties and dopamine (DA) detection capability of SU-8 photoresist based pyrolytic carbon (PyC) as well as its biocompatibility with neural cells. This approach is compatible with microfabrication techniques which is crucial for device development. X-ray photoelectron spectroscopy shows that PyC consists 98.5% of carbon, while oxygen plasma treatment (PyC-O2) increases the amount of oxygen up to 27.1%. PyC showed nearly reversible (ΔEp 63 mV) electron transfer kinetics towards outer sphere redox probe (Ru(NH3)62+/3+), while the reaction on PyC-O2 was quasi-reversible (ΔEp > 75 mV). DA showed both diffusion and adsorption-defined reaction kinetics with fast electron transfer with the ΔEp values of 50 mV and 30 mV, for PyC and PyC-O2, respectively. The strong interaction between the hydroxyl groups on the surface and DA, as confirmed by simulations, facilitates the redox reactions of DA. DA showed a linear response in the measured physiologically relevant range (50 nM–1 μM) and sensitivities were 1.2 A M−1 cm−2 for PyC and 2.7 A M−1 cm−2 for PyC-O2. Plasma oxidation (PyC-O2) improved cell adhesion even more than poly-L-lysine (PLL) coating on PyC, but best adhesion was achieved on PLL coated PyC-O2. Glial cells, neuroblastoma cells and neural stem cells all showed similar behavior.

A micropillar array electrospray ionization (μPESI) platform fabricated from thiol–enes with 56 individual polyethylene glycol coated μPESI chips for bioanalytical mass spectrometry is introduced. Bioanalysis capability is shown by measurement of a protein, a protein digest and a cell lysate sample. The thiol–ene polyethylene glycol (PEG) coated μPESI chip allows the use of a wide range of aqueous–organic solvent compositions and provides a detection limit at 60 zeptomole level (6 × 10−20 mol) for a peptide standard.

Tunable and responsive surfaces offer routes to multiple functionalities ranging from superhydrophobic surfaces to controlled adhesion. Inspired by cilia structure in the respiratory pathway, magnetically responsive periodic arrays of flexible and magnetic thiol-ene micropillars are fabricated. Omnidirectional collective bending of the pillar array in magnetic field is shown. Local non-contact actuation of a single pillar is achieved using an electromagnetic needle to probe the responsiveness and the elastic properties of the pillars by comparing the effect of thiol-ene crosslinking density to pillar bending. The suitable thiol-ene components for flexible and stiff magnetic micropillars and the workable range of thiol-to-allyl ratio are identified. The wettability of the magnetic pillars can be tailored by chemical and topography modification of the pillar surface. Low-surface-energy self-assembled monolayers are grafted by UV-assisted surface activation, which is also used for surface topography modification by covalent bonding of micro- and nanoparticles to the pillar surface. The modified thiol-ene micopillars are resistant to capillarity-driven collapse and they exhibit low contact angle hysteresis, allowing water droplet motion driven by repeated bending and recovery of the magnetic pillars in an external magnetic field. Transport of polyethylene microspheres is also demonstrated.

Organically modified ceramic polymers (ORMOCERs) have attracted substantial interest in biomicrofluidic applications owing to their inherent biocompatibility and high optical transparency even in the near-ultraviolet (UV) range. However, the processes for metallization of ORMOCERs as well as for sealing of metallized surfaces have not been fully developed. In this study, we developed metallization processes for a commercial ORMOCER formulation, Ormocomp, covering several commonly used metals, including aluminum, silver, gold, and platinum. The obtained metallizations were systematically characterized with respect to adhesion (with and without adhesion layers), resistivity, and stability during use (in electrochemical assays). In addition to metal adhesion, the possibility for Ormocomp bonding over each metal as well as sufficient step coverage to guarantee conductivity over topographical features (e.g., over microchannel edges) was addressed with a view to the implementation of not only planar, but also three-dimensional on-chip sensing elements. The feasibility of the developed metallization for implementation of microfluidic electrochemical assays was demonstrated by fabricating an electrophoresis separation chip, compatible with a commercial bipotentiostat, and incorporating integrated working, reference, and auxiliary electrodes for amperometric detection of an electrochemically active pharmaceutical, acetaminophen.

Pattern formation of pyrolyzed carbon (PyC) and tetrahedral amorphous carbon (ta-C) thin films were investigated at micro- and nanoscale. Micro- and nanopillars were fabricated from both materials, and their biocompatibility was studied with cell viability tests. Carbon materials are known to be very challenging to pattern. Here we demonstrate two approaches to create biocompatible carbon features. The microtopographies were 2 &mu, m or 20 &mu, m pillars (1:1 aspect ratio) with three different pillar layouts (square-grid, hexa-grid, or random-grid orientation). The nanoscale topography consisted of random nanopillars fabricated by maskless anisotropic etching. The PyC structures were fabricated with photolithography and embossing techniques in SU-8 photopolymer which was pyrolyzed in an inert atmosphere. The ta-C is a thin film coating, and the structures for it were fabricated on silicon substrates. Despite different fabrication methods, both materials were formed into comparable micro- and nanostructures. Mouse neural stem cells were cultured on the samples (without any coatings) and their viability was evaluated with colorimetric viability assay. All samples expressed good biocompatibility, but the topography has only a minor effect on viability. Two &mu, m pillars in ta-C shows increased cell count and aggregation compared to planar ta-C reference sample. The presented materials and fabrication techniques are well suited for applications that require carbon chemistry and benefit from large surface area and topography, such as electrophysiological and -chemical sensors for in vivo and in vitro measurements.

The low fabrication cost of SU-8-based devices has opened the fields of point-of-care devices (POC), µTAS and Lab-on-Chip technologies, which call for cheap and disposable devices. Often this translates to free-standing, suspended devices and a reusable carrier wafer. This necessitates a sacrificial layer to release the devices from the substrates. Both inorganic (metals and oxides) and organic materials (polymers) have been used as sacrificial materials, but they fall short for fabrication and releasing multilayer SU-8 devices. We propose photoresist AZ 15nXT (MicroChemicals GmbH, Ulm, Germany) to be used as a sacrificial layer. AZ 15nXT is stable during SU-8 processing, making it suitable for fabricating free-standing multilayer devices. We show two methods for cross-linking AZ 15nXT for stable sacrificial layers and three routes for sacrificial release of the multilayer SU-8 devices. We demonstrate the capability of our release processes by fabrication of a three-layer free-standing microfluidic electrospray ionization (ESI) chip and a free-standing multilayer device with electrodes in a microchannel.

Prior to sensory experience spontaneous activity appears to play a fundamental role in the correct formation of prominent functional features of different cortical regions. The use of anaesthesia during pregnancy such as ketamine is largely considered to negatively affect neuronal development by interfering with synaptic transmission. Interestingly, the characteristics of spontaneous activity as well as the acute functional effects of maternal anaesthesia remain largely untested in the embryonic cortex in vivo. In the present work, we performed in vivo imaging of spontaneous calcium activity and cell motility in the marginal zone of the cortex of E14-15 embryos connected to the mother. We made use of a preparation where the blood circulation from the mother through the umbilical cord is preserved and fluctuations in intracellular calcium in the embryonic frontal cortex are acquired using two-photon imaging. We found that spontaneous transients were either sporadic or correlated in clusters of neuronal ensembles at this age. These events were not sensitive to maternal isoflurane anaesthesia but were strongly inhibited by acute in situ or maternal application of low concentration of the anaesthetic ketamine (a non-competitive antagonist of NMDA receptors). Moreover, simultaneous imaging of cell motility revealed a correlated strong sensitivity to ketamine. These results show that anaesthetic compounds can differ significantly in their impact on spontaneous early cortical activity as well as motility of cells in the marginal zone. The effects found in this study may be relevant in the etiology of heightened vulnerability to cerebral dysfunction associated with the use of ketamine during pregnancy.

UDP-glucuronosyltransferases (UGTs), located in the endoplasmic reticulum of liver cells, are an important family of enzymes, responsible for the biotransformation of several endogenous and exogenous chemicals, including therapeutic drugs. However, the phenomenon of 'latency', i.e., full UGT activity revealed by disruption of the microsomal membrane, poses substantial challenges for predicting drug clearance based on in vitro glucuronidation assays. This work introduces a microfluidic reactor design comprising immobilized human liver microsomes to facilitate the study of UGT-mediated drug clearance under flow-through conditions. The performance of the microreactor is characterized using glucuronidation of 8-hydroxyquinoline (via multiple UGTs) and zidovudine (via UGT2B7) as the model reactions. With the help of alamethicin and albumin effects, we show that conducting UGT metabolism assays under flow conditions facilitates in-depth mechanistic studies, which may also shed light on UGT latency.

Synaptic adhesion molecules are key components in development of the brain, and in the formation of neuronal circuits, as they are central in the assembly and maturation of chemical synapses. Several families of neuronal adhesion molecules have been identified such as the neuronal cell adhesion molecules, neurexins and neuroligins, and in particular recently several leucine-rich repeat proteins, e.g., Netrin G-ligands, SLITRKs, and LRRTMs. The LRRTMs form a family of four proteins. They have been implicated in excitatory glutamatergic synapse function and were specifically characterized as ligands for neurexins in excitatory synapse formation and maintenance. In addition, LRRTM3 and LRRTM4 have been found to be ligands for heparan sulfate proteoglycans, including glypican. We report here the crystal structure of a thermostabilized mouse LRRTM2, with a Tm 30 °C higher than that of the wild-type protein. We localized the neurexin binding site to the concave surface based on protein engineering, sequence conservation, and prior information about the interaction of the ligand with neurexins, which allowed us to propose a tentative model for the LRRTM-neurexin interaction complex. We also determined affinities of the thermostabilized LRRTM2 and wild-type LRRTM1 and LRRTM2 for neurexin-β1 with and without Ca(2+). Cell culture studies and binding experiments show that the engineered protein is functional and capable of forming synapselike contacts. The structural and functional data presented here provide the first structure of an LRRTM protein and allow us to propose a model for the molecular mechanism of LRRTM function in the synaptic adhesion.

Microfabrication technology has been used to prepare a microchip sensor-array with six sets of platinum electrodes. Chromium/platinum (10 nm/100 nm thick) were sputtered on a borosilicate wafer and patterned by wet etching method. The electrodes were designed with working electrode area of 700×400 μm in the middle and a 200 μm wide and 2600 μm long counter electrode surrounding it from three sides in a U-shape. The connection pads (1000×1500 μm) were located at the edge of a sensor-array chip. Silicon wafer was etched through to form holes with slanting side walls for immobilization cavities. The silicon and the borosilicate wafers were adhesion bonded with SU-8 epoxy resin. The cyclic voltammetry and electrochemical impedance experiments were carried out in a three-electrode electrochemical system to characterize the fabricated sensor-array chip. The results show that the current density depends on the electrode potential sweep rate ν. Also, current density depends on the concentration of potassium hexacyanoferrate(III). At slow potential sweep rates (ν≤0.01 V s−1) the steady-state signal is achieved and the electrodes behave as micro-electrodes. Such an array is a promising candidate for fast and simple biochemical oxygen demand (BOD) measurements.

Directional imbibition of oils (hexadecane, tetradecane, and dodecane) and water is demonstrated on a chemically patterned silicon micropillar array. Four different directional imbibition types are shown: unidirectional, two types of bidirectional and tridirectional imbibition. The surfaces consist of a silicon micropillar array with an overlaid surface chemistry pattern. This configuration leads to anisotropic wetting behaviour into various directions of the advancing meniscus. Due to the free energy landscape obtained, the advancing meniscus gets pinned in some directions (determined by the surface chemistry pattern) while it is free to move to the remaining directions. The conditions for directional imbibition and design criteria for the surfaces are derived and discussed.