Microflow sensing and control using an in-channel birefringent biomembrane.

This study describes the function, optimization, and demonstration of a new class of passive, low-cost microfluidic flow meters based on birefringent chitosan biomembranes analyzed by polarized microscopy. We subjected the membrane to dynamic flow conditions while monitoring the real-time response of its optical properties. We obtained figures of merit, including the linear response operating range (0 to 65 µL min-1), minimum response time (250 ms), sensitivity (2.03% × 10-3 µL-1 min), and minimum sensor longevity (1 week). In addition, possible sources of interference were identified. Finally, we demonstrate the membrane as a low-cost flow rate measurement device for the close loop control of a commercial pressure-driven pump. Preliminary experiments using a basic PID controller with the membrane-based flow rate measurement device showed that stable control could be achieved and the system could reach steady-state behavior in less than 15 seconds. Analysis of fundamental limits to sensor response time indicate the potential for faster steady-state behaviour.

Microfluidic membraneless microbial fuel cells: new protocols for record power densities.

The main hurdle in leveraging microfluidic advantages in membraneless MFCs is their low electrode area-normalized power. For nearly a decade, maximum power densities have remained stagnant, while at the same time macrosystems continue to gather pace. To bridge this growing gap, we showcase a strategy that focuses on (i) technology improvements, (ii) establishment of record areal power densities, and (iii) presentation of different normalization methods that complement areal power densities and enable direct comparisons across all MFC scales. Using a pure-culture Geobacter sulfurreducens electroactive biofilm (EAB) in a new membraneless MFC that adheres to the strategy above, we observed optimal anode colonization, resulting in the highest recorded electrode areal power density for a microfluidic MFC of 3.88 W m-2 (24.37 kW m-3). We also consider new power normalization methods that may be more appropriate for comparison to other works. Normalized by the wetted cross-section area between electrodes accounts for constraints in electrode/electrolyte contact, resulting in power densities as high as 8.08 W m-2. Alternatively, we present a method to normalize by the flow rate to account for acetate supply, obtaining normalized energy recovery values of 0.025 kW h m-3. With these results, the performance gap between micro- and macroscale MFCs is closed, and a road map to move forward is presented.

A spectIR-fluidic reactor for monitoring fast chemical reaction kinetics with on-chip attenuated total reflection Fourier transform infrared spectroscopy.

Microfluidics has emerged as a powerful technology with diverse applications in microbiology, medicine, chemistry, and physics. While its potential for controlling and studying chemical reactions is well recognized, the extraction and analysis of useful chemical information generated within microfluidic devices remain challenging. This is mainly due to the limited tools available for in situ measurements of chemical reactions. In this study, we present a proof-of-concept spectIR-fluidic reactor design that combines microfluidics with Fourier transform infrared (FTIR) spectroscopy for in situ kinetic studies of fast reactions. By integrating a multi-ridge silicon attenuated total reflection (ATR) wafer into the microfluidic device, we enable multi-point measurements for precise reaction time monitoring. As such, this work establishes a validated foundation for studying fast chemical reactions using on-chip ATR-FTIR spectroscopy in a microfluidic reactor environment, which enables simultaneous monitoring of reagents, intermediates, and products using a phosphate proton transfer reaction. The spectIR-fluidic reactor platform offers customizable designs, allowing for the investigation of reactions with various time scales, and has the potential to significantly advance studies exploring reaction mechanisms and optimization.

SpectIR-fluidics: completely customizable microfluidic cartridges for high sensitivity on-chip infrared spectroscopy with point-of-application studies on bacterial biofilms.

We present a generalizable fabrication method for a new class of analytical devices that merges virtually any microfluidic design with high-sensitivity on-chip attenuated total reflection (ATR) sampling using any standard Fourier transform infrared (FTIR) spectrometer. Termed "spectIR-fluidics", a major design feature is the integration of a multi-groove silicon ATR crystal into a microfluidic device, compared with previous approaches in which the ATR surface served as a structural support for the entire device. This was accomplished by the design, fabrication, and aligned bonding of a highly engineered ATR sensing layer, which con```tains a seamlessly embedded ATR crystal on the channel side and an optical access port that matched the spectrometer light path characteristics at the device exterior. The refocused role of the ATR crystal as a dedicated analytical element, combined with optimized light coupling to the spectrometer, results in limits of detection as low as 540 nM for a D-glucose solution, arbitrarily complex channel features that are fully enclosed, and up to 18 world-to-chip connections. Three purpose-built spectIR-fluidic cartridges are used in a series of validation experiments followed by several point-of-application studies on biofilms from the gut microbiota of plastic-consuming insects using a small portable spectrometer.

Portable impedance-sensing device for microorganism characterization in the field.

A variety of biosensors have been proposed to quickly detect and measure the properties of individual microorganisms among heterogeneous populations, but challenges related to cost, portability, stability, sensitivity, and power consumption limit their applicability. This study proposes a portable microfluidic device based on impedance flow-cytometry and electrical impedance spectroscopy that can detect and quantify the size of microparticles larger than 45 µm, such as algae and microplastics. The system is low cost ($300), portable (5 cm [Formula: see text] 5 cm), low-power (1.2 W), and easily fabricated utilizing a 3D-printer and industrial printed circuit board technology. The main novelty we demonstrate is the use of square wave excitation signal for impedance measurements with quadrature phase-sensitive detectors. A linked algorithm removes the errors associated to higher order harmonics. After validating the performance of the device for complex impedance models, we used it to detect and differentiate between polyethylene microbeads of sizes between 63 and 83 µm, and buccal cells between 45 and 70 µm. A precision of 3% is reported for the measured impedance and a minimum size of 45 µm is reported for the particle characterization.

Recent progress in microbial fuel cells using substrates from diverse sources.

Increasing untreated environmental outputs from industry and the rising human population have increased the burden of wastewater and other waste streams on the environment. The most prevalent wastewater treatment methods include the activated sludge process, which requires aeration and is, therefore, energy and cost-intensive. The current trend towards a circular economy facilitates the recovery of waste materials as a resource. Along with the amount, the complexity of wastewater is increasing day by day. Therefore, wastewater treatment processes must be transformed into cost-effective and sustainable methods. Microbial fuel cells (MFCs) use electroactive microbes to extract chemical energy from waste organic molecules to generate electricity via waste treatment. This review focuses use of MFCs as an energy converter using wastewater from various sources. The different substrate sources that are evaluated include industrial, agricultural, domestic, and pharmaceutical types. The article also highlights the effect of operational parameters such as organic load, pH, current, and concentration on the MFC output. The article also covers MFC functioning with respect to the substrate, and the associated performance parameters, such as power generation and wastewater treatment matrices, are given. The review also illustrates the success stories of various MFC configurations. We emphasize the significant measures required to fill in the gaps related to the effect of substrate type on different MFC configurations, identification of microbes for use as biocatalysts, and development of biocathodes for the further improvement of the system. Finally, we shortlisted the best performing substrates based on the maximum current and power, Coulombic efficiency, and chemical oxygen demand removal upon the treatment of substrates in MFCs. This information will guide industries that wish to use MFC technology to treat generated effluent from various processes.

Mainstreaming microfluidic microbial fuel cells: a biocompatible membrane grown in situ improves performance and versatility.

A recent trend in microfluidic microbial fuel cells (MFCs) is to exclude a separation membrane, instead, relying on the physics of laminar flow to maintain isolation between anode and cathode compartments. To avoid solution crossover, the electrodes may be separated by distances of several millimeters, but this negatively affects the internal resistance and undermines a prime advantage of microscale MFCs. Therefore, we propose a facile method for in situ synthesis of a micromembrane that supports sub-millimeter electrode spacing. Membrane synthesis in situ reduces device fabrication complexity, and the proposed design avoids electrode contamination during its synthesis. Comparing results to a state-of-the-art membraneless MFC with 6 mm inter-electrode distances, the sub-millimeter membrane MFC under comparable flow conditions had an internal resistance that was 60% lower, power and current densities that were respectively 45% and 290% higher, and acetate conversion efficiencies that were 8 times higher. The enhanced flow stability provided stable operation under imbalanced flow conditions and delivered continuous increases to power density of up to 30% for flow rate increases of 100 times over baseline levels. As a result, maximum outputs obtained were 660 mW m-1 and 3.5 A m-1. These are the highest reported for microfluidic MFCs using pure culture bacteria, which advances the goal of competing with mainstream MFC formats.

Scanning Aperture Approach for Spatially Selective ATR-FTIR Spectroscopy: Application to Microfluidics.

We present a novel spectroscopy accessory that can easily convert any Fourier transform infrared (FTIR) spectrometer into a fully automated mapping and assaying system. The accessory uses a multiridge attenuated total reflection (ATR) wafer as the sensing element coupled with a moving aperture that is used to select the regions of interest on the wafer. In this demonstration, the accessory is combined with a series of parallel micropatterned channels, which are positioned co-linear with the light-coupling ridges on the opposite side of the ATR wafer. The ATR spectroscopy microfluidic assay accessory (ASMAA) was used in continuous mapping mode to scan perpendicular to the ATR ridges, revealing complex but repeatable oscillations in the spectral intensities. To understand this behavior, the light path through the optical components was simulated with consideration of the aperture position, ridge-to-channel alignment, and excitation beam profile. With this approach, the simulation reproduced the experimental mapping results and provided evidence that the measurement position and area changed with the aperture position. To demonstrate the assay mode, we obtained spectra along the centerline of individual microchannels and determined noise baselines and limits of detection.

Versatile Microfluidic Platform for Automated Live-Cell Hyperspectral Imaging Applied to Cold Climate Cyanobacterial Biofilms.

Microfluidic bioanalytical platforms are driving discoveries from synthetic biology to the health sciences. In this work, we present a platform for in vivo live-cell imaging and automated species detection in mixed cyanobacterial biofilms from cold climate environments. Using a multimodal microscope with custom optics applied to a chip with six parallel growth channels, we monitored biofilm dynamics via continuous imaging at natural irradiance levels. Machine learning algorithms were applied to the collected hyperspectral images for automatic segmentation of mixed-species biofilms into individual species of cyanobacteria with similar filamentous morphology. The coupling of microfluidic technology with modern multimodal imaging and computer vision systems provides a versatile platform for the study of cause-and-effect scenarios of cyanobacterial biofilms, which are important elements of many ecosystems, including lakes and rivers of the polar regions.

A microfluidic approach to micromembrane synthesis for complex release profiles of nanocarriers.

Physically crosslinked microscale biomembranes synthesized from pure chitosan are designed and demonstrated for pH-triggered release of embedded functionalized mesoporous silica nanoparticles. Nanoparticle-loaded membranes are formed in a microfluidic channel at the junction between accurately controlled co-flowing streams to achieve highly tuneable membrane properties. After formation, the loaded membranes remain stable until contact with physiological acidic conditions, resulting in controlled nanoparticle release. Furthermore, nanoparticle-loaded membranes with complex layered architectures are synthesized using different flow schemes, thus enabling customized nanoparticle release profiles. These novel materials are well-suited for integration within small medical devices as well as off-chip applications.

On the nature of "skeletal" biofilm patterns, "hidden" heterogeneity and the role of bubbles to reveal them.

A short communication on the recent paper by Jang et al. discusses the role of "mushroom" structures and effects of nearly static bubbles on nascent biofilms.

Microfluidic bioanalytical flow cells for biofilm studies: a review.

Bacterial biofilms are among the oldest and most prevalent multicellular life forms on Earth and are increasingly relevant in research areas related to industrial fouling, medicine and biotechnology. The main hurdles to obtaining definitive experimental results include time-varying biofilm properties, structural and chemical heterogeneity, and especially their strong sensitivity to environmental cues. Therefore, in addition to judicious choice of measurement tools, a well-designed biofilm study requires strict control over experimental conditions, more so than most chemical studies. Due to excellent control over a host of physiochemical parameters, microfluidic flow cells have become indispensable in microbiological studies. Not surprisingly, the number of lab-on-chip studies focusing on biofilms and other microbiological systems with expanded analytical capabilities has expanded rapidly in the past decade. In this paper, we comprehensively review the current state of microfluidic bioanalytical research applied to bacterial biofilms and offer a perspective on new approaches that are expected to drive continued advances in this field.

Linear Scanning ATR-FTIR for Chemical Mapping and High-Throughput Studies of Pseudomonas sp. Biofilms in Microfluidic Channels.

A fully automated linear scanning attenuated total reflection (ATR) accessory is presented for Fourier transform infrared (FTIR) spectroscopy. The approach is based on the accurate displacement of a multibounce ATR crystal relative to a stationary infrared beam. To ensure accurate positioning and to provide a second sample characterization mode, a custom-built microscope was integrated into the system and the computerized work flow. Custom software includes automated control and measurement routines with a straightforward user interface for selecting parameters and monitoring experimental progress. This cost-effective modular system can be implemented on any research-grade spectrometer with a standard sample compartment for new bioanalytical chemistry studies. The system was validated and optimized for use with microfluidic flow cells containing growing Pseudomonas sp. bacterial biofilms. The complementarity among the scan positioning accuracy, measurement spatial resolution, and the microchannel dimensions paves the way for parallel biological assays with real-time control over environmental parameters and minimal manual labor. By rotating the channel orientation relative to the beam path, the system could also be used for acquisition of linear biochemical maps and stitched microscope images along the channel length.

A new look at bubbles during biofilm inoculation reveals pronounced effects on growth and patterning.

Specially designed microfluidic bioflow cells were used to temporarily trap microbubbles during different inoculation stages of Pseudomonas sp. biofilms. Despite being eliminated many hours before biofilm appearance, templated growth could occur at former bubble positions. Bubble-templated growth was either continuous or in ring patterns, depending on the stage of inoculation when the bubbles were introduced. Templated biofilms were strongly enhanced in terms of their growth kinetics and structural homogeneity. High resolution confocal imaging showed two separate bubble-induced bacterial trapping modes, which were responsible for the altered biofilm development. It is concluded that static bubbles can be exploited for fundamental improvements to bioreactor performance, as well as open new avenues to study isolated bacteria and small colonies.

Recent Advancements towards Full-System Microfluidics.

Microfluidics is quickly becoming a key technology in an expanding range of fields, such as medical sciences, biosensing, bioactuation, chemical synthesis, and more. This is helping its transformation from a promising R&D tool to commercially viable technology. Fuelling this expansion is the intensified focus on automation and enhanced functionality through integration of complex electrical control, mechanical properties, in situ sensing and flow control. Here we highlight recent contributions to the Sensors Special Issue series called "Microfluidics-Based Microsystem Integration Research" under the following categories: (i) Device fabrication to support complex functionality; (ii) New methods for flow control and mixing; (iii) Towards routine analysis and point of care applications; (iv) In situ characterization; and (v) Plug and play microfluidics.

Hydrodynamic Effects on Biofilms at the Biointerface Using a Microfluidic Electrochemical Cell: Case Study of Pseudomonas sp.

The anchoring biofilm layer is expected to exhibit a different response to environmental stresses than for portions in the bulk, due to the protection from other strata and the proximity to the attachment surface. The effect of hydrodynamic stress on surface-adhered biofilm layers was tested using a specially designed microfluidic bio flow cell with an embedded three-electrode detection system. In situ electrochemical impedance spectroscopy (EIS) measurements of biocapacitance and bioresistance of Pseudomonas sp. biofilms were conducted during the growth phase and under different shear flow conditions with verification by other surface sensitive techniques. Distinct, but reversible changes to the amount of biofilm and its structure at the attachment surface were observed during the application of elevated shear stress. In contrast, regular microscopy revealed permanent distortion to the biofilm bulk, in the form of streamers and ripples. Following the application of extreme shear stresses, complete removal of significant portions of biofilm outer layers occurred, but this did not change the measured quantity of biofilm at the electrode attachment surface. The structure of the remaining biofilm, however, appeared to be modified and susceptible to further changes following application of shear stress directly to the unprotected biofilm layers at the attachment surface.

One-Step Fabrication of Microchannels with Integrated Three Dimensional Features by Hot Intrusion Embossing.

We build on the concept of hot intrusion embossing to develop a one-step fabrication method for thermoplastic microfluidic channels containing integrated three-dimensional features. This was accomplished with simple, rapid-to-fabricate imprint templates containing microcavities that locally control the intrusion of heated thermoplastic based on their cross-sectional geometries. The use of circular, rectangular and triangular cavity geometries was demonstrated for the purposes of forming posts, multi-focal length microlense arrays, walls, steps, tapered features and three-dimensional serpentine microchannels. Process variables, such as temperature and pressure, controlled feature dimensions without affecting the overall microchannel geometry. The approach was demonstrated for polycarbonate, cycloolefin copolymer and polystyrene, but in principle is applicable to any thermoplastic. The approach is a step forward towards rapid fabrication of complex, robust, microfluidic platforms with integrated multi-functional elements.

Towards a Multifunctional Electrochemical Sensing and Niosome Generation Lab-on-Chip Platform Based on a Plug-and-Play Concept.

In this paper, we present a new modular lab on a chip design for multimodal neurotransmitter (NT) sensing and niosome generation based on a plug-and-play concept. This architecture is a first step toward an automated platform for an automated modulation of neurotransmitter concentration to understand and/or treat neurodegenerative diseases. A modular approach has been adopted in order to handle measurement or drug delivery or both measurement and drug delivery simultaneously. The system is composed of three fully independent modules: three-channel peristaltic micropumping system, a three-channel potentiostat and a multi-unit microfluidic system composed of pseudo-Y and cross-shape channels containing a miniature electrode array. The system was wirelessly controlled by a computer interface. The system is compact, with all the microfluidic and sensing components packaged in a 5 cm × 4 cm × 4 cm box. Applied to serotonin, a linear calibration curve down to 0.125 mM, with a limit of detection of 31 µ M was collected at unfunctionalized electrodes. Added sensitivity and selectivity was achieved by incorporating functionalized electrodes for dopamine sensing. Electrode functionalization was achieved with gold nanoparticles and using DNA and o-phenylene diamine polymer. The as-configured platform is demonstrated as a central component toward an "intelligent" drug delivery system based on a feedback loop to monitor drug delivery.

Electrochemical imaging for microfluidics: a full-system approach.

Electrochemistry is developed as a new chemical imaging modality for microfluidics. The technique is based on multipoint voltammetry using an embedded 20 × 10 miniature electrode array implemented on a customized printed circuit board. Electrode durability was enhanced by chemical modification of the electrode surfaces, which enabled continuous, stable use for over 2 months. A system-level approach enables automatic calibration, data acquisition and data processing through a graphical user interface. Following data processing, redox currents and peak positions are extracted from location-specific voltammograms and converted into pixels of an "electrochemical image". The system is validated by imaging steady-state and dynamic laminar flow patterns of flow-confined solutions of the redox pairs Fe(CN)6(3-/4-) or multi-redox environments that include coflowing Ru(NH3)6(2+/3+) solutions. The images obtained are compared with flow simulations and optical images for validation. A strategy to achieve measurements with spatial resolution smaller than the individual electrodes is also demonstrated as an avenue to enhance image spatial resolution. It is expected that this new approach to chemical imaging will expand the applicability of microfluidics in certain areas of chemistry and biology without requiring expertise in electrochemistry.

Live-streaming: Time-lapse video evidence of novel streamer formation mechanism and varying viscosity.

Time-lapse videos of growing biofilms were analyzed using a background subtraction method, which removed camouflaging effects from the heterogeneous field of view to reveal evidence of streamer formation from optically dense biofilm segments. In addition, quantitative measurements of biofilm velocity and optical density, combined with mathematical modeling, demonstrated that streamer formation occurred from mature, high-viscosity biofilms. We propose a streamer formation mechanism by sudden partial detachment, as opposed to continuous elongation as observed in other microfluidic studies. Additionally, streamer formation occurred in straight microchannels, as opposed to serpentine or pseudo-porous channels, as previously reported.