Chemical Concentration Gradients Research Articles

Micro-/nanofluidic device for tunable generation of a concentration gradient: application to Caenorhabditis elegans chemotaxis

In this study, we propose a novel micro-/nanofluidic device that can generate a chemical concentration gradient using a parallel nanochannel as gradient generator. This device is easy to fabricate, showing high reproducibility. Its main feature is the multiple-nanochannel-based gradient generator, which permits the diffusion of small molecules and tunably generates concentration gradients. The nanopattern for the nanochannels can be rapidly and easily fabricated by wrinkling a diamond-like carbon thin film which is deposited on a polydimethylsiloxane substrate; the generation of the concentration gradient can be adjusted by controlling the dimensions of the nanochannels. The developed gradient generator is embedded into a microfluidic device to study chemotaxis in the nematode Caenorhabditis elegans, which has a highly developed chemosensory system and can detect a wide variety of chemical molecules. This device shows good performance for rapid analysis of C. elegans chemotaxis under sodium chloride stimuli.

Adjustable, rapidly switching microfluidic gradient generation using focused travelling surface acoustic waves

We demonstrate a simple device to generate chemical concentration gradients in a microfluidic channel using focused travelling surface acoustic waves (F-TSAW). A pair of curved interdigitated metal electrodes deposited on the surface of a piezoelectric (LiNbO3) substrate disseminate high frequency sound waves when actuated by an alternating current source. The F-TSAW produces chaotic acoustic streaming flow upon its interaction with the fluid inside a microfluidic channel, which mixes confluent streams of chemicals in a controlled fashion for an adjustable and rapidly switching gradient generation.

Fabrication of Unique Chemical Patterns and Concentration Gradients with Visible Light

A modular and general method based on a photomediated ATRA reaction for the spatially controlled functionalization of surfaces with visible light is reported. The ability to control reactivity with light intensity combined with the orthogonality of ATRA chemistry allows well-defined chemically differentiated monolayers and complex nonlinear chemical concentration gradients to be easily prepared. Use of light to mediate these reactions permits spatial regulation and the generation of unique, multifunctional chemical gradients.

Microfluidic flow-free generation of chemical concentration gradients

This paper presents a class of novel microfluidic concentration gradient generation (CGG) devices that create temporally stable chemical concentration gradients with complex shapes in a flow-free environment. The devices feature a two-layer channel design and the incorporation of a semipermeable membrane, which effectively segregates the concentration gradient region in the lower layer from the flow of reagent sample (simply “sample” onward) and buffer in the upper layer. In the mean time, free diffusion across the membrane constantly replenishes sample and buffer to maintain a stable concentration. The shapes of the concentration gradients are controlled by the geometries of the micro-channels and chambers. Concentration gradients with complex shapes can be achieved by piecewise combining constituent gradients with elementary shapes. Capable of generating concentration gradients in a flow-free environment, our devices eliminate undesirable flow stimulation on biological cells under investigation, while maintaining a stable chemical environment for cell studies.

Motility analysis of bacteria‐based microrobot (bacteriobot) using chemical gradient microchamber

A bacteria-based microrobot (bacteriobot) was proposed and investigated as a new type of active drug delivery system because of its useful advantages, such as active tumor targeting, bacteria-mediated tumor diagnosis, and therapy. In this study, we fabricated a bacteriobot with enhanced motility by selective attachment of flagellar bacteria (Salmonella typhimurium). Through selective bovine serum albumin (BSA) pattering on hydrophobic polystyrene (PS) microbeads, many S. typhimurium could be selectively attached only on the unpatterned surface of PS microbead. For the evaluation of the chemotactic motility of the bacteriobot, we developed a microfluidic chamber which can generate a stable concentration gradient of bacterial chemotactic chemicals. Prior to the evaluation of the bacteriobot, we first evaluated the directional chemotactic motility of S. typhimurium using the proposed microfluidic chamber, which contained a bacterial chemo-attractant (L-aspartic acid) and a chemo-repellent (NiSO4 ), respectively. Compared to density of the control group in the microfluidic chamber without any chemical gradient, S. typhimurium increased by about 16% in the L-aspartic acid gradient region and decreased by about 22% in the NiSO4 gradient region. Second, we evaluated the bacteriobot's directional motility by using this microfluidic chamber. The chemotactic directional motility of the bacteriobot increased by 14% and decreased by 13% in the concentration gradients of L-aspartic acid and NiSO4 , respectively. These results confirm that the bacteriobot with selectively patterned S. typhimurium shows chemotaxis motility very similar to that of S. typhimurium. Moreover, the directional motilities of the bacteria and bacteriobot could be demonstrated quantitatively through the proposed microfluidic chamber.

The role of multiple chemotactic mechanisms in a model of chemotaxis in C. elegans: different mechanisms are specialised for different environments

Unlike simpler organisms, C. elegans possesses several distinct chemosensory pathways and chemotactic mechanisms. These mechanisms and pathways are individually capable of driving chemotaxis in a chemical concentration gradient. However, it is not understood if they are redundant or co-operate in more sophisticated ways. Here we examine the specialisation of different chemotactic mechanisms in a model of chemotaxis to NaCl. We explore the performance of different chemotactic mechanisms in a range of chemical gradients and show that, in the model, far from being redundant, the mechanisms are specialised both for different environments and for distinct features within those environments. We also show that the chemotactic drive mediated by the ASE pathway is not robust to the presence of noise in the chemical gradient. This problem cannot be solved along the ASE pathway without destroying its ability to drive chemotaxis. Instead, we show that robustness to noise can be achieved by introducing a second, much slower NaCl-sensing pathway. This secondary pathway is simpler than the ASE pathway, in the sense that it can respond to either up-steps or down-steps in NaCl but not both, and could correspond to one of several candidates in the literature which we identify and evaluate. This work provides one possible explanation of why there are multiple NaCl sensing pathways and chemotactic mechanisms in C. elegans: rather than being redundant the different pathways and mechanism are specialised both for the characteristics of different environments and for distinct features within a single environment.

Rapid Detection of Drugs of Abuse in Saliva Using Surface Enhanced Raman Spectroscopy and Microfluidics

We present a microfluidic device that detects trace concentrations of drugs of abuse in saliva within minutes using surface-enhanced Raman spectroscopy (SERS). Its operation is demonstrated using methamphetamine. The detection scheme exploits concentration gradients of chemicals, fostered by the laminar flow in the device, to control the interactions between the analyte, silver nanoparticles (Ag-NPs), and a salt. Also, since all species interact while advecting downstream, the relevant reaction coordinates occur with respect to the position in the channel. The system was designed to allow the analyte first to diffuse into the side stream containing the Ag-NPs, on which it is allowed to adsorb, before salt ions are introduced, causing the Ag-NPs to aggregate, and so creating species with strong SERS signal. The device allows partial separation via diffusion of the analyte from the complex mixture. Also, the reproducible salt-induced NP aggregation decouples the aggregation reaction (necessary for strong SERS) from the analyte concentration or charge. This method enables the creation of a region where detection of the analyte of interest via SERS is optimal, and dramatically extends the classes of molecules and quality of signals that can be measured using SERS, compared to bulk solution methods. The spatial distribution of the SERS signals was used to map the degree of nanoparticle aggregation and species diffusion in the channel, which, together with numerical simulations, was used to describe the kinetics of the colloid aggregation reaction, and to determine the optimal location in the channel for SERS interrogation.

Towards a dynamic clamp for neuro-chemical modalities

The classic dynamic clamp technique uses a real-time electrical interface between living cells and neural simulations in order to investigate hypotheses about neural function and structure [3]. However, it has two major drawback [2]: the electrodes can clamp electrically only a small section of a cell, and hence frequently do not control the behavior of whole cells; and all control to-date has been concentrated on the electric properties of neurons, neglecting their chemical state. As noted in [2], the latter has been done by necessity, since until recently registering or controlling the chemical state of either neurons or the chemical environment in which they reside has been essentially impossible with the level of precision needed to simulate the appropriate dynamics of the various extracellular chemical players. In this manuscript we present an expansion of the dynamic clamp method to include simulation and control of the effects of a variety of chemicals associated with neural signaling. To achieve that, we use the emergent discipline of microfluidics [4], which deals specifically with the behavior, precise control and manipulation of fluids at the microscale. We use a novel combination of microfluidic and nanosensor technology to add sensing and control of chemical concentrations to the dynamic clamp technique. Specifically, we use a microfluidic chip to generate distinct chemical concentration gradients (ions or neuromodulators), register the concentrations with embedded nanosensors, and use the processed signals as an input to simulations of a neural cell. The ultimate goal of this project is to close the loop, and provide control signals to the microfluidic lab to mimic the interaction of the simulated cell with other cells in its chemical environment, by modifying the chemical concentrations in the microfluidic lab environment to reflect simulated outputs of the model neurons. Here we used Hodgkin-Huxley type neurons, CdVdt=Io+IHH+ICawith added calcium-gated currents, ICa=m∞(V)2hPmaxz2F2RTV([Ca]in-[Ca]oute-zVFRT1-e-zVFRT) In order to predict the distribution of specific chemicals inside a microfluidic chamber, 2-D Computational Fluid Dynamics technique was used. In this study, the commercial software FLUENT® 6.3 was used to build the computational domain, the models for a microfluidic mixer and a neuron chamber by using finite element methods. Figure 1 Shows the simulated microfluidic device (A) and simulated square pulse flow at pts C1 and C2 (B). The responses of the model neuron to changes in Ca++ concentration are shown in panel C. The device in (A) was realized physically, using standard lithography ...

Fabricating Nanoscale Chemical Gradients with ThermoChemical NanoLithography

Production of chemical concentration gradients on the submicrometer scale remains a formidable challenge, despite the broad range of potential applications and their ubiquity throughout nature. We present a strategy to quantitatively prescribe spatial variations in functional group concentration using ThermoChemical NanoLithography (TCNL). The approach uses a heated cantilever to drive a localized nanoscale chemical reaction at an interface, where a reactant is transformed into a product. We show using friction force microscopy that localized gradients in the product concentration have a spatial resolution of ~20 nm where the entire concentration profile is confined to sub-180 nm. To gain quantitative control over the concentration, we introduce a chemical kinetics model of the thermally driven nanoreaction that shows excellent agreement with experiments. The comparison provides a calibration of the nonlinear dependence of product concentration versus temperature, which we use to design two-dimensional temperature maps encoding the prescription for linear and nonlinear gradients. The resultant chemical nanopatterns show high fidelity to the user-defined patterns, including the ability to realize complex chemical patterns with arbitrary variations in peak concentration with a spatial resolution of 180 nm or better. While this work focuses on producing chemical gradients of amine groups, other functionalities are a straightforward modification. We envision that using the basic scheme introduced here, quantitative TCNL will be capable of patterning gradients of other exploitable physical or chemical properties such as fluorescence in conjugated polymers and conductivity in graphene. The access to submicrometer chemical concentration and gradient patterning provides a new dimension of control for nanolithography.

Cellulose-based hydrogels with excellent microstructural replication ability and cytocompatibility for microfluidic devices

In the face of challenges in the development of excellent biocompatible materials for microfluidic device fabrication, we demonstrated that cross-linked cellulose (RCC) hydrogel can be used as the bulk material for microchips. The cellulose hydrogel was prepared from cellulose solution dissolved in an 8 wt% LiOH/15 wt% urea aqueous system with cooling by crosslinking with epichlorohydrin. Collagen as a key extracellular matrix component for promoting cell cultivation was cross-linked in the cellulose hydrogel to obtain cellulose–collagen (RCC/C) hybrid hydrogels. The experimental results revealed that cellulose-based hydrogel microchips with well-defined 2D or 3D microstructures possessed excellent structural replication ability, good mechanical properties, and cytocompatibility for cell culture as well as excellent dimensional stability at elevated temperature. The hydrogel, as a transparent microchip material, had no effect on the fluorescence behaviors of FITC-dextran and rhodamine-dextran, leading to the good conjunction with fluorescent detection and imaging. Moreover, collagen could be immobilized in the RCC/C hydrogel scaffold for promoting cell growth and generating stable chemical concentration gradients, leading to superior cytocompatibility. This work provides new hydrogel materials for the microfluidic technology field and mimicks a 3D cell culture microenvironment for cell-based tissue engineering and drug screening.

Pattern Formation and Cross-Diffusion for a Chemotaxis Model

Chemotaxis is the feature movement of cell or an organism along a chemical concentration gradient. The mathematical analysis of chemotaxis modelss how a plenitude of spatial patterns such as the chemotaxis models applied to skin pigmentation patterns, that lead to aggregations of one type of pigment cells into a striped spatial pattern. The analysis of pattern formation can be traced to a seminal paper by Turing [1], who established that an action-diffusion system can generate stable nonuniform patterns in space if the components of the system interact with each other.Our motivation is the numerical simulations of the pattern formation for a volume-filling chemotaxis model. In [2], the effect of volume-filling is expressed through a nonlinear squeezing probability. We investigate pattern formation using Turing's principle and the standard argument used by Murray [3,4]. Next, we introduce an implicit nite volume scheme; it is presented on a general mesh satisfying the orthogonality condition [5,6].The originality of this scheme is the upstream approach to discretize thecross-diffusion term. Finally, we present some numerical results showing the spatial patterns for the chemotaxis model.

Modelling of the physiological response of the brain to ischaemic stroke

Identification of salvageable brain tissue is a major challenge when planning the treatment of ischaemic stroke. As the standard technique used in this context, the perfusion-diffusion mismatch, has not shown total accuracy, there is an ongoing search for new imaging protocols that could better identify the region of the brain at risk and for new physiological models that could, on the one hand, incorporate the imaged parameters and predict the evolution of the condition for the individual, and, on the other hand, identify future biomarkers and thus suggest new directions for the design of imaging protocols. Recently, models of cellular metabolism after stroke and blood-brain barrier transport at tissue level have been introduced. We now extend these results by developing a model of the propagation of key metabolites in the brain's extracellular space owing to stroke-related oedema and chemical concentration gradients between the ischaemic and normal brain. We also couple the resulting chemical changes in the extracellular space with cellular metabolism. Our work enables the first patient-specific simulations of stroke progression with finite volume models to be made.

Construction of oxygen and chemical concentration gradients in a single microfluidic device for studying tumor cell–drug interactions in a dynamic hypoxia microenvironment

Recent microfluidic advancements in oxygen gradients have greatly promoted controllable oxygen-sensitive cellular investigations at microscale resolution. However, multi-gradient integration in a single microfluidic device for tissue-mimicking cell investigation is not yet well established. In this study, we describe a method that can generate oxygen and chemical concentration gradients in a single microfluidic device via the formation of an oxygen gradient in a chamber and a chemical concentration gradient between adjacent chambers. The oxygen gradient dynamics were systematically investigated, and were quantitatively controlled using simple exchange between the aerial oxygen and the oxygen-free conditions in the gas-permeable polydimethylsiloxane channel. Meanwhile, the chemical gradient dynamics was generated using a special channel-branched device. For potential medical applications of the established oxygen and chemical concentration gradients, a tumor cell therapy assessment was performed using two antitumor drugs (tirapazamine and bleomycin) and two tumor cell lines (human lung adenocarcinoma A549 cells and human cervical carcinoma HeLa cells). The results of the proof-of-concept experiment indicate the dose-dependent antitumor effect of the drugs and hypoxia-induced cytotoxicity of tirapazamine. We demonstrate that the integration of oxygen and chemical concentration gradients in a single device can be applied to investigating oxygen- and chemical-sensitive cell events, which can also be valuable in the development of multi-gradient generating procedures and specific drug screening.

Gas/liquid sensing via chemotaxis of Euglena cells confined in an isolated micro-aquarium

We demonstrate on-chip gas/liquid sensing by using the chemotaxis of live bacteria (Euglena gracilis) confined in an isolated micro-aquarium, and gas/liquid permeation through porous polydimethylsiloxane (PDMS). The sensing chip consisted of one closed micro-aquarium and two separated bypass microchannels along the perimeter of the micro-aquarium. Test gas/liquid and reference samples were introduced into the two individual microchannels separately, and the gas/liquid permeated through the PDMS walls and dissolved in the micro-aquarium water, resulting in a chemical concentration gradient in the micro-aquarium. By employing the closed micro-aquarium isolated from sample flows, we succeeded in measuring the chemotaxis of Euglena for a gas substance quantitatively, which cannot be achieved with the conventional flow-type or hydro-gel-type microfluidic devices. We found positive (negative) chemotaxis for CO2 concentrations below (above) 15%, with 64 ppm as the minimum concentration affecting the cells. We also observed chemotaxis for ethanol and H2O2. By supplying culture medium via the microchannels, the Euglena culture remained alive for more than 2 months. The sensing chip is thus useful for culturing cells and using them for environmental toxicity/nutrition studies by monitoring their motion.

Preface to Special Topic: Microfluidics in Cancer Research

Preface to Special Topic: Microfluidics in Cancer Research

Effect of solutal Marangoni convection on motion, coarsening, and coalescence of droplets in a monotectic system

In this paper, we study the effect of solutal Marangoni convection (SMC) on the microstructure evolution in a monotectic system, using the convective Cahn-Hilliard and Navier-Stokes equations with a capillary tensor contributed by the chemical concentration gradient. At first, we simulate the spontaneous motion of two distant droplets induced by SMC and compare our results with an analytical solution. We then compute the coalescence of two droplets in contact and coarsening of two distant droplets considering different sizes. We further study the influence of SMC on the evolution of phase separation processes inside the spinodal region for Fe-50 at %Sn and Fe-40 at %Sn alloys. In the former case, we rationalize our results using Fourier spectra and in the latter case, we compare the size distribution of droplets with the LSW theory.

Self-similar dynamics of bacterial chemotaxis

Colonies of bacteria grown on thin agar plate exhibit fractal patterns as a result of adaptation to their environments. The bacterial colony pattern formation is regulated crucially by chemotaxis, the movement of cells along a chemical concentration gradient. Here, the dynamics of pattern formation in a bacterial colony is investigated theoretically through a continuum model that considers chemotaxis. In the case of the gradient sensed by the bacterium is nearly uniform, the bacterial colony patterns are self-similar, which means they look the same at every scale. The scaling law of the bacterial colony growth has been revealed explicitly. Chemotaxis biases the movement of the bacterial population in colony to trend toward the chemical attractant. Moreover, the bacterial colonies evolve for a long time as the traveling wave with a sharp front.

Golem geographies

In our response, below, we undertake a deeper engagement with one of the authors, Elizabeth Grosz, that a number of the respondents have evoked, implicitly as well as explicitly, in their response to our text and, in particular, Grosz's arguments on art as of the chaotic and of the animal. Whilst our comments on the bowerbird speak to the artistry involved in a sexuated mode of reproduction that increases biological difference, we emphasise an abyssal moment in our initial text wherein we move from a consideration of sexual selection to symbiosis and accretion as productive of diversity, terms that no longer sit quite so comfortably in the lexicon of the ‘animal’. Rather, we implicate aesthetics and an artistry across and down chemical concentration gradients.

Microstructural evolution and high temperature failure of ferritic to austenitic dissimilar welds

Dissimilar metal welds between ferritic and austenitic alloys are used extensively in power plants. Premature failure of such welds can occur below the expected creep life of either base metal. This article reviews microstructural evolution in dissimilar welds and describes factors that contribute to premature failure. The microstructure in the as welded condition consists of a sharp chemical concentration gradient across the fusion line. Martensite forms within this gradient due to high hardenability and rapid cooling rates from welding. Upon aging, carbon diffuses down the chemical potential gradient from the ferritic steel toward the austenitic alloy. This can lead to formation of a soft carbon denuded zone in the ferritic steel, and nucleation and growth of carbides in the austenitic steel that produce high hardness. These differences in microstructure and hardness occur over distances of about 50–100 μm. Failure is attributed to the steep microstructural and mechanical property gradients, the large difference in coefficient of thermal expansion, formation of interfacial carbides that promote creep cavity formation, and preferential oxidation of the ferritic steel. Information is also provided on available creep rupture properties, remaining life estimation techniques, current best practices and research in progress.

Active Brownian motion tunable by light

Active Brownian particles are capable of taking up energy from their environment and converting it into directed motion; examples range from chemotactic cells and bacteria to artificial micro-swimmers. We have recently demonstrated that Janus particles, i.e. gold-capped colloidal spheres, suspended in a critical binary liquid mixture perform active Brownian motion when illuminated by light. In this paper, we investigate in more detail their swimming mechanism, leading to active Brownian motion. We show that the illumination-borne heating induces a local asymmetric demixing of the binary mixture, generating a spatial chemical concentration gradient which is responsible for the particle’s self-diffusiophoretic motion. We study this effect as a function of the functionalization of the gold cap, the particle size and the illumination intensity: the functionalization determines what component of the binary mixture is preferentially adsorbed at the cap and the swimming direction (towards or away from the cap); the particle size determines the rotational diffusion and, therefore, the random reorientation of the particle; and the intensity tunes the strength of the heating and, therefore, of the motion. Finally, we harness this dependence of the swimming strength on the illumination intensity to investigate the behavior of a micro-swimmer in a spatial light gradient, where its swimming properties are space-dependent.