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Multispecies microbial communities drive most ecosystems on Earth. Chemical and biological interactions within these communities can affect the survival of individual members and the entire community. However, the prohibitively high number of possible interactions within a microbial community has made the characterization of factors that influence community development challenging. Here, we report a Microbial Community Interaction (μCI) device to advance the systematic study of chemical and biological interactions within a microbial community. The μCI creates a combinatorial landscape made up of an array of triangular wells interconnected with circular wells, which each contains either a different chemical or microbial strain, generating chemical gradients and revealing biological interactions. Bacillus cereus UW85 containing green fluorescent protein provided the "target" readout in the triangular wells, and antibiotics or microorganisms in adjacent circular wells are designated the "variables." The μCI device revealed that gentamicin and vancomycin are antagonistic to each other in inhibiting the target B. cereus UW85, displaying weaker inhibitory activity when used in combination than alone. We identified three-member communities constructed with isolates from the plant rhizosphere that increased or decreased the growth of B. cereus. The μCI device enables both strain-level and community-level insight. The scalable geometric design of the μCI device enables experiments with high combinatorial efficiency, thereby providing a simple, scalable platform for systematic interrogation of three-factor interactions that influence microorganisms in solitary or community life. [ABSTRACT FROM AUTHOR]

As a leading cause of mortality and morbidity, stroke constitutes a significant global health burden. Ischemic stroke accounts for 80% of cases and occurs due to an arterial thrombus, which impedes cerebral blood flow and rapidly leads to cell death. As the most abundant cell type within the central nervous system, astrocytes play a critical role within the injured brain. We developed a novel microphysiological platform that permits the induction of spatiotemporally controlled nutrient gradients, allowing us to study astrocytic response during and after transient nutrient deprivation. Within 24 h of inducing starvation in the platform, nutrient deprivation led to multiple changes in astrocyte response, from metabolic perturbations to gene expression changes, and cell viability. Furthermore, we observed that nutrient restoration did not reverse the functional changes in astrocyte metabolism, which mirrors reperfusion injury observed in vivo. We also identified alterations in numerous glucose metabolism-associated genes, many of which remained upregulated or downregulated even after restoration of the nutrient supply. Together, these findings suggest that astrocyte activation during and after nutrient starvation induces plastic changes that may underpin persistent stroke-induced functional impairment. Overall, our innovative device presents interesting potential to be used in the development of new therapies to improve tissue repair and even cognitive recovery after stroke. [ABSTRACT FROM AUTHOR]

Enzymes are important catalysts in biochemical reactions with superior regio, stereo, and substrate selectivity. However, enzymatic reaction systems have drawbacks including product inhibition, difficulty recycling, and poor stability. Importantly, the rate of an enzyme catalyzed reaction diminishes rapidly due to product inhibition and substrate depletion, making it difficult for many enzymes to catalyze a reaction to completion. The outcome is a mixture of unreacted substrates being present in the final reaction, necessitating additional separation steps that increase costs. This study presents a microfluidic reactor for accelerating enzyme catalyzed reactions using a countercurrent design that continuously removes products and adds fresh substrate into the reaction, allowing enzymes to operate under better reaction conditions. It demonstrates that countercurrent flow accelerates enzymatic reactions in our system up to 36 % for horseradish peroxidase and 21 % for β‐glucosidase compared to cocurrent flow, and the resulting reaction solution contains highly pure product with minimal substrate contamination. [ABSTRACT FROM AUTHOR]

The addition of reagents for assays in digital microfluidic (DMF) systems is traditionally done by merging of droplets containing different analytes or reagents in solution. However, this process significantly increases droplet volume after each step, resulting in dilution of the analyte and reagents. Here, we report a new technique for performing reagent additions to aqueous droplets without significantly increasing the droplet's volume: volume-less reagent delivery (VRD). VRD is enabled by a physical phenomenon we call "exclusive liquid repellency" (ELR), which describes an aqueous/oil/solid 3-phase system where the aqueous phase can be fully repelled from a solid phase (contact angle ∼180°). When performing VRD, a reagent of interest in solution is deposited onto the ELR solid surface and allowed to dry. The ELR surface containing the dried reagent is then immersed under oil, followed by introduction of an aqueous droplet. By dragging the aqueous droplet over the spot of dried reagent using paramagnetic particles or via a physical sliding wall, the droplet can then recover and reconstitute the reagent with negligible increase in its total volume, returning the ELR surface to its initial liquid repellent state in the process. We demonstrate that VRD can be performed across a wide range of reagent types including sugars, proteins (antibodies), nucleic acids (DNA), antibiotics, and even complex enzyme/substrate/buffer "kit" mixtures. We believe VRD is a flexible and powerful technique which can further the development of self-contained, multi-step assays in DMF and other microfluidic systems. [ABSTRACT FROM AUTHOR]

Double-exclusive liquid repellency (double-ELR) is an extreme wettability phenomenon in which adjacent regions selectively and completely repel immiscible liquids with different surface chemistries on a non-textured substrate (i.e., a substrate in absence of micro/nano-structures). Under double-ELR conditions, each liquid exhibits no physical contact (contact angle of 180°) with its non-preferred surface chemistry, thus enabling complete partitioning of adjacent fluidic volumes (e.g., between water and oil). This enables a new type of cell culture-based assay, where cell loss from common failure modes (e.g., biofouling from inadvertent cell adhesion, detrimental moisture loss/gain, and liquid handling dead volumes) is significantly mitigated. Importantly, the principles of double-ELR were leveraged to achieve underoil sweep patterning, a no-loss, robust and high-throughput distribution of sub-microliter volumes of aqueous media (and cells). In addition to high-efficiency distribution via sweep patterning, double-ELR can be used to construct “modular” (i.e., easily implemented and/or linked together with spatial and temporal control) higher-order architectures for in vitro imitation of physiologically relevant microenvironments that are of particular interest within the cell assay community, including multi-phenotype cultures with excellent spatial and temporal control, three-dimensional layered multi-phenotype cultures, cultures with selective mechanical cues of extracellular matrix (i.e., collagen fiber alignment), and spheroid cultures. Together, these features of double-ELR uniquely facilitate culture and high content analysis of limited cellular samples (e.g., a few hundred to a few thousand cells). [ABSTRACT FROM AUTHOR]

Here, we report the novel application of a material with self-concentrating properties for enhancing the sensitivity of immunoassays. Termed as glass microbubbles, they are antibody functionalized buoyant hollow glass microspheres that simultaneously float and concentrate into a dense monolayer when dispensed in a liquid droplet. This self-concentrating charactaristic of the microbubbles allow for autonomous signal localization, which translates to a higher sensitivity compared to other microparticle-based immunoassays. We then demonstrated a “microbubble array” platform consisting of the glass microbubbles floating in a microfluidic liquid hemisphere array for performing multiplex immunoassays. [ABSTRACT FROM AUTHOR]

In vitro cell motility assays are frequently used in the study of cell migration in response to anti-cancer drug treatment. Microfluidic systems represent a unique tool for the in vitro analysis of cell motility. However, they usually rely on using time-lapse microscopy to record the spatial temporal locations of the individual cells being tested. This has created a bottleneck for microfluidic systems to perform high-throughput experiments due to requirement of a costly time-lapse microscopy system. Here, we describe the development of a portable microfluidic device for endpoint analysis of cell motility. The reported device incorporates a cell alignment feature to position the seeded cells on the same initial location, so that the cells' motilities can be analyzed based on their locations at the end of the experiment after the cells have migrated. We show that the device was able to assess cancer cell motility after treatment with a migration inhibitory drug Indole-3-carbinol on MDA-MB-231 breast cancer cells, demonstrating the applicability of our device in screening anti-cancer drug compounds on cancer cells. [ABSTRACT FROM PUBLISHER]