Supervised machine learning in microfluidic impedance flow cytometry for improved particle size determination.

The assessment of particle and cell size in electrical microfluidic flow cytometers has become common practice. Nevertheless, in flow cytometers with coplanar electrodes accurate determination of particle size is difficult, owing to the inhomogeneous electric field. Pre-defined signal templates and compensation methods have been introduced to correct for this positional dependence, but are cumbersome when dealing with irregular signal shapes. We introduce a simple and accurate post-processing method without the use of pre-defined signal templates and compensation functions using supervised machine learning. We implemented a multiple linear regression model and show an average reduction of the particle diameter variation by 37% with respect to an earlier processing method based on a feature extraction algorithm and compensation function. Furthermore, we demonstrate its application in flow cytometry by determining the size distribution of a population of small (4.6 ± 0.9 µm) and large (5.9 ± 0.8 µm) yeast cells. The improved performance of this coplanar, two electrode chip enables precise cell size determination in easy to fabricate impedance flow cytometers.

Self-Propelled Detachment upon Coalescence of Surface Bubbles.

The removal of microbubbles from substrates is crucial for the efficiency of many catalytic and electrochemical gas evolution reactions in liquids. The current work investigates the coalescence and detachment of bubbles generated from catalytic decomposition of hydrogen peroxide. Self-propelled detachment, induced by the coalescence of two bubbles, is observed at sizes much smaller than those determined by buoyancy. Upon coalescence, the released surface energy is partly dissipated by the bubble oscillations, working against viscous drag. The remaining energy is converted to the kinetic energy of the out-of-plane jumping motion of the merged bubble. The critical ratio of the parent bubble sizes for the jumping to occur is theoretically derived from an energy balance argument and found to be in agreement with the experimental results. The present results provide both physical insight for the bubble interactions and practical strategies for applications in chemical engineering and renewable energy technologies like electrolysis.

Highly parallelized human embryonic stem cell differentiation to cardiac mesoderm in nanoliter chambers on a microfluidic chip.

Human stem cell-derived cells and tissues hold considerable potential for applications in regenerative medicine, disease modeling and drug discovery. The generation, culture and differentiation of stem cells in low-volume, automated and parallelized microfluidic chips hold great promise to accelerate the research in this domain. Here, we show that we can differentiate human embryonic stem cells (hESCs) to early cardiac mesodermal cells in microfluidic chambers that have a volume of only 30 nanoliters, using discontinuous medium perfusion. 64 of these chambers were parallelized on a chip which contained integrated valves to spatiotemporally isolate the chambers and automate cell culture medium exchanges. To confirm cell pluripotency, we tracked hESC proliferation and immunostained the cells for pluripotency markers SOX2 and OCT3/4. During differentiation, we investigated the effect of different medium perfusion frequencies on cell reorganization and the expression of the early cardiac mesoderm reporter MESP1mCherry by live-cell imaging. Our study demonstrates that microfluidic technology can be used to automatically culture, differentiate and study hESC in very low-volume culture chambers even without continuous medium perfusion. This result is an important step towards further automation and parallelization in stem cell technology.

Single catalyst particle diagnostics in a microreactor for performing multiphase hydrogenation reactions.

Since inter- and intra-particle heterogeneities in catalyst particles are more the rule than the exception, it is advantageous to perform high-throughput screening for the activity of single catalyst particles. A multiphase system (gas/liquid/solid) is developed, where droplet-based microfluidics and optical detection are combined for the analysis of single catalyst particles by safely performing a hydrogenation study on in-house synthesized hollow Pd/SiO2 catalyst microparticles, in a polydimethylsiloxane (PDMS) microreactor. A two-phase segmented flow system of particle-containing droplets is combined with a parallel gas-reactant channel separated from the flow channel by a 50 µm thick gas permeable PDMS wall. In this paper, the developed microreactor system is showcased by monitoring the Pd-catalyzed hydrogenation of methylene blue. A discoloration of blue to brown visualizes the hydrogenation activity happening in a high-throughput fashion on the single Pd/SiO2 spherical catalyst microparticles, which are encapsulated in 50 nL-sized droplets. By measuring the reagent concentration at various spots along the length of the channel the reaction time can be determined, which is proportional to the residence time in the channel. The developed experimental platform opens new possibilities for single catalyst particle diagnostics in a multiphase environment.

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.

Monitoring Contractile Cardiomyocytes via Impedance Using Multipurpose Thin Film Ruthenium Oxide Electrodes.

A ruthenium oxide (RuOx) electrode was used to monitor contractile events of human pluripotent stem cells-derived cardiomyocytes (hPSC-CMs) through electrical impedance spectroscopy (EIS). Using RuOx electrodes presents an advantage over standard thin film Pt electrodes because the RuOx electrodes can also be used as electrochemical sensor for pH, O2, and nitric oxide, providing multisensory functionality with the same electrode. First, the EIS signal was validated in an optically transparent well-plate setup using Pt wire electrodes. This way, visual data could be recorded simultaneously. Frequency analyses of both EIS and the visual data revealed almost identical frequency components. This suggests both the EIS and visual data captured the similar events of the beating of (an area of) hPSC-CMs. Similar EIS measurement was then performed using the RuOx electrode, which yielded comparable signal and periodicity. This mode of operation adds to the versatility of the RuOx electrode's use in in vitro studies.

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.

Electrochemical Control of pH in Nanoliter Volumes.

The electrochemical management of the proton concentration in miniaturized dimensions opens the way to control and parallelize multistep chemical reactions, but still it faces many challenges linked to the efficient proton generation and control of their diffusion. Here we present a device operated electrochemically that demonstrates the control of the pH in a cell of ∼140 nL. The device comprises a microfluidic reactor integrated with a pneumatic mechanism that allows the exchange of reagents and the isolation of protons to decrease the effect of their diffusion. We monitored the pH with a fluorescence marker and calculated the final value from the redox currents. We demonstrate a large pH amplitude control from neutral pH values beyond the fluorescence marker range at pH 5. On the basis of the calculations from the Faradaic currents, the minimum pH reached should undergo pH ∼ 0.9. The pH contrast between neutral and acid pH cells can be maintained during periods longer than 15 min with an appropriate design of a diffusion barrier.

Integrated microfluidic biosensing platform for simultaneous confocal microscopy and electrophysiological measurements on bilayer lipid membranes and ion channels.

Combining high-resolution imaging and electrophysiological recordings is key for various types of experimentation on lipid bilayers and ion channels. Here, we propose an integrated biosensing platform consisting of a microfluidic cartridge and a dedicated chip-holder to conduct such dual measurements on suspended lipid bilayers, in a user-friendly manner. To illustrate the potential of the integrated platform, we characterize lipid bilayers in terms of thickness and fluidity while simultaneously monitoring single ion channel currents. For that purpose, POPC lipid bilayers are supplemented with a fluorescently-tagged phospholipid (NBD-PE, 1% mol) for Fluorescence Recovery After Photobleaching (FRAP) measurements and a model ion channel (gramicidin, 1 nM). These combined measurements reveal that NBD-PE has no effect on the lipid bilayer thickness while gramicidin induces thinning of the membrane. Furthermore, the presence of gramicidin does not alter the lipid bilayer fluidity. Surprisingly, in lipid bilayers supplemented with both probes, a reduction in gramicidin open probability and lifetime is observed compared to lipid bilayers with gramicidin only, suggesting an influence of NBD-PE on the gramicidin ion function. Altogether, our proposed microfluidic biosensing platform in combination with the herein presented multi-parametric measurement scheme paves the way to explore the interdependent relationship between lipid bilayer properties and ion channel function.

Eccentricity scale independence for scene perception in the first tens of milliseconds.

Visual processing of scenes in the first tens of milliseconds relies on global image summary statistics rather than localized processing. Although natural scenes typically involve our entire visual field, scenes are usually presented experimentally at limited eccentricity. Receptive-field size increases with foveal eccentricity while increasingly pooling activity from local receptive fields. Here, we asked to what extent an observer's performance on a scene-gist perception task depends on the contents of the scene as well as on the eccentricity of the scene. We manipulated the scene content by applying window and scotoma masks. In addition, we changed presentation eccentricity independent of image content by upscaling and downscaling the scenes. We find that discrimination is strongly affected when the scene is presented with a window of 5°, showing only the central part rather than the whole scene. Performance is, however, eccentricity scale independent provided that the same scene content is presented and a comparable area of the surface of primary visual cortex is activated. We furthermore show that this eccentricity scale independence holds for shorter presentation times, down to 17 ms in some scene-discrimination tasks, but not for the naturalness-discrimination task.

Human V2A: A map of the peripheral visual hemifield with functional connections to scene-selective cortex.

Humans can recognize a scene in the blink of an eye. This gist-based visual scene perception is thought to be underpinned by specialized visual processing emphasizing the visual periphery at a cortical locus relatively low in the visual processing hierarchy. Using wide-field retinotopic mapping and population receptive field (pRF) modeling, we identified a new visual hemifield map anterior of area V2d and inferior to area V6, which we propose to call area V2A. Based on its location relative to other visual areas, V2A may correspond to area 23V described in nonhuman primates. The pRF analysis revealed unique receptive field properties for V2A: a large (FWHM ∼23°) and constant receptive field size across the central ∼70° of the visual field. Resting-state fMRI connectivity analysis further suggests that V2A is ideally suited to quickly feed the scene-processing network with information that is not biased towards the center of the visual field. Our findings not only indicate a likely cortical locus for the initial stages of gist-based visual scene perception, but also suggest a reappraisal of the organization of human dorsomedial occipital cortex with a strip of separate hemifield representations anterior to the early visual areas (V1, V2d, and V3d).

Ruthenium Oxide Nanorods as Potentiometric pH Sensor for Organs-On-Chip Purposes.

A ruthenium oxide (RuOx) sensor for potentiometric pH sensing is currently being developed for organs-on-chip purposes. The sensor was fabricated from a Ru(OH)3 precursor, resulting in RuOx nanorods after heating. An open-circuit potential of the RuOx electrode showed a near-Nernstian response of -58.05 mV/pH, with good selectivity against potentially interfering ions (lithium, sulfate, chloride, and calcium ions). The preconditioned electrode (stored in liquid) had a long-term drift of -0.8 mV/h, and its response rate was less than 2 s. Sensitivity to oxygen was observed at an order of magnitude lower than other reported metal-oxide pH sensors. Together with miniaturizability, the RuOx pH sensor proves to be a suitable pH sensor for organs-on-chip studies.

Lab-on-a-Chip: Frontier Science in the Classroom.

Lab-on-a-chip technology is brought into the classroom through development of a lesson series with hands-on practicals. Students can discover the principles of microfluidics with different practicals covering laminar flow, micromixing, and droplet generation, as well as trapping and counting beads. A quite affordable novel production technique using scissor-cut and laser-cut lamination sheets is presented, which provides good insight into how scientific lab-on-a-chip devices are produced. In this way high school students can now produce lab-on-a-chip devices using lamination sheets and their own lab-on-a-chip design. We begin with a review of previous reports on the use of lab-on-a-chip technology in classrooms, followed by an overview of the practicals and projects we have developed with student safety in mind. We conclude with an educational scenario and some initial promising results for student learning outcomes.

Magnetophoretic Sorting of Single Catalyst Particles.

A better understanding of the deactivation processes taking place within solid catalysts is vital to design better ones. However, since inter-particle heterogeneities are more a rule than an exception, particle sorting is crucial to analyse single catalyst particles in detail. Microfluidics offers new possibilities to sort catalysts at the single particle level. Herein, we report a first-of-its-kind 3D printed magnetophoretic chip able to sort catalyst particles by their magnetic moment. Fluid catalytic cracking (FCC) particles were separated based on their Fe content. Magnetophoretic sorting shows that large Fe aggregates exist within 20 % of the FCC particles with the highest Fe content. The availability of Brønsted acid sites decreases with increasing Fe content. This work paves the way towards a high-throughput catalyst diagnostics platform to determine why specific catalyst particles perform better than others.

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.

A meta-analysis of perceptual and cognitive functions involved in useful-field-of-view test performance.

The useful-field-of-view (UFOV) test measures the amount of information someone can extract from a visual scene in one glance. Its scores show relatively strong relationships with everyday activities. The UFOV test consists of three computer tests, suggested to measure processing speed and central vision, divided attention, and selective attention. However, other functions seem to be involved as well. In order to investigate the contribution of these suggested and other perceptual and cognitive functions, we performed a meta-analysis of 116 Pearson's correlation coefficients between UFOV scores and other test scores reported in 18 peer-reviewed articles. We divided these correlations into nine domains: attention, executive functioning, general cognition, memory, spatial ability, visual closure, contrast sensitivity, visual processing speed, and visual acuity. A multivariate mixed-effects model analysis revealed that each domain correlated significantly with each of the UFOV subtest scores. These correlations were stronger for Subtests 2 and 3 than for Subtest 1. Furthermore, some domains were more strongly correlated to the UFOV than others across subtests. We did not find interaction effects between subtest and domain, indicating that none of the UFOV subtests is more selectively sensitive to a particular domain than the others. Thus, none of the three UFOV subtests seem to measure one clear construct. Instead, a range of visual and cognitive functions is involved. Perhaps this is the reason for the UFOV's high ecological validity, as it involves many functions at once, making it harder to compensate if one of them fails.

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.