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Microhabitat conditions determine the magnitude and speed of microbial processes but have been challenging to investigate. In this study we used microfluidic devices to determine the effect of the spatial distortion of a pore space on fungal and bacterial growth, interactions, and substrate degradation. The devices contained channels differing in bending angles and order. Sharper angles reduced fungal and bacterial biomass, especially when angles were repeated in the same direction. Substrate degradation was only decreased by sharper angles when fungi and bacteria were grown together. Investigation at the cellular scale suggests that this was caused by fungal habitat modification, since hyphae branched in sharp and repeated turns, blocking the dispersal of bacteria and the substrate. Our results demonstrate how the geometry of microstructures can influence microbial activity. This can be transferable to soil pore spaces, where spatial occlusion and microbial feedback on microstructures is thought to explain organic matter stabilization., Arellano Caicedo et al. use microfluidic devices to determine how spatial distortion of pore spaces in soil organic matter affects bacterial and fungal growth and substrate degradation. Their newly conceptualized chip design allows the quantification of bacterial and fungal biomass, and ecosystem function organic matter cycling with help of fluorescent markers in different microstructures.

Handling of submicron-sized objects is important in many biochemical and biomedical applications, but few methods today can precisely manipulate this range of particles. We present gradient acoustic focusing that enables flow-through particle separation of submicron particles and cells and we apply it for separation of bacteria from blood lysate to facilitate their detection in whole blood for improved diagnostics. To control suspended objects below the classical 2µm size limit for acoustic focusing, we introduce a co-flowing acoustic impedance gradient to generate a stabilizing acoustic volume force that supresses acoustic streaming. The method is validated theoretically and experimentally using polystyrene particles, Staphylococcus aureus, Streptococcus pneumoniae and Escherichia coli. The applicability of the method is demonstrated by the separation of bacteria from selectively chemically lysed blood. Combined with downstream operations, this new approach opens up for novel methods for sepsis diagnostics.

A finite-element model is presented for numerical simulation in three dimensions of acoustophoresis of suspended microparticles in a microchannel embedded in a polymer chip and driven by an attached piezoelectric transducer at MHz frequencies. In accordance with the recently introduced principle of whole-system ultrasound resonances, an optimal resonance mode is identified that is related to an acoustic resonance of the combined transducer-chip-channel system and not to the conventional pressure half-wave resonance of the microchannel. The acoustophoretic action in the microchannel is of comparable quality and strength to conventional silicon-glass or pure glass devices. The numerical predictions are validated by acoustic focusing experiments on 5-um-diameter polystyrene particles suspended inside a microchannel, which was milled into a PMMA-chip. The system was driven anti-symmetrically by a piezoelectric transducer, driven by a 30-V peak-to-peak AC-voltage in the range from 0.5 to 2.5 MHz, leading to acoustic energy densities of 13 J/m^3 and particle focusing times of 6.6 s., 12 pages, 7 pdf figures, pdf-latex

Microfluidic soil chips render optical access to the naturally opaque soil systems and enable direct investigation of microbial growth and interactions in micro-structurally and chemically controlled environments. However, chemical analyses of these interactions at high spatial and temporal resolution are still lacking. Here we propose that the use of advanced microspectroscopy techniques, namely infrared absorption, Raman scattering and synchrotron radiation based X-ray microspectroscopy, in microfluidic soil chips would make it possible to approach these phenomena. They allow monitoring biogeochemical processes in and around soil microbial cells growing in the reproducibly designed microenvironments within the chips at (sub)micrometer scale. Complementary use of several of the microspectroscopy techniques is beneficial for obtaining information about both molecular and elemental composition, oxidation states and local structure of the elements in the sample. Ultimately, we argue that microspectroscopy in microfluidic chips can lead to relevant breakthroughs in frontier research areas in soil science, such as (1) analysis of chemical responses of microbes to environmental triggers at micro-scale spatial resolution, (2) phenotypical identification and phylogenetic classification of single cells of soil microbes in situ, (3) determining spatially and time resolved effects of heavy metals and organic pollutants, including microplastics, on soils and (4) spatially resolved analysis of soil organic matter dynamics for better understanding of soil carbon storage. Tailoring the chip design to achieve optical transparency to the radiation type used by the different microspectroscopy methods is crucial to achieve this; therefore, we expect that this perspective will inspire the scientific community to use the proposed approaches and thus push both the technical development of the microspectroscopy suitable soil chips and the research frontier in soil science.

Effective separation methods for fractionating blood components are needed for numerous diagnostic and research applications. This paper presents the use of acoustophoresis, an ultrasound based microfluidic separation technology, for label-free, gentle and continuous separation of mononuclear cells (MNCs) from diluted whole blood. Red blood cells (RBCs) and MNCs behave similar in an acoustic standing wave field, compromising acoustic separation of MNC from RBC in standard buffer systems. However, by optimizing the buffer conditions and thereby changing the acoustophoretic mobility of the cells, we were able to enrich MNCs relative to RBCs by a factor of 2,800 with MNC recoveries up to 88%. The acoustophoretic microchip can perform cell separation at a processing rate of more than 1 × 105 cells/s, corresponding to 5 µl/min undiluted whole blood equivalent. Thus, acoustophoresis can be easily integrated with further down-stream applications such as flow cytometry, making it a superior alternative to existing MNC isolation techniques.

Soil is likely the most complex ecosystem on earth. Despite the global importance and extraordinary diversity of soils, they have been notoriously challenging to study. We show how pioneering microfluidic techniques provide new ways of studying soil microbial ecology by allowing simulation and manipulation of chemical conditions and physical structures at the microscale in soil model habitats.

This paper describes an integrated microsystem for rapid separation, enrichment, and detection of bacteria from blood, addressing the unmet clinical need for rapid sepsis diagnostics. The blood sample is first processed in an acoustophoresis chip, where red blood cells are focused to the center of the channel by an acoustic standing wave and sequentially removed. The bacteria-containing plasma proceeds to a glass capillary with a localized acoustic standing wave field where the bacteria are trapped onto suspended polystyrene particles. The trapped bacteria are subsequently washed while held in the acoustic trap and released into a polymer microchip containing dried polymerase chain reaction (PCR) reagents, followed by thermocycling for target sequence amplification. The entire process is completed in less than 2 h. Testing with Pseudomonas putida spiked into whole blood revealed a detection limit of 1000 bacteria/mL for this first-generation analysis system. In samples from septic patients, the system was able to detect Escherichia coli in half of the cases identified by blood culture. This indicates that the current system detects bacteria in patient samples in the upper part of the of clinically relevant bacteria concentration range and that a further developed acoustic sample preparation system may open the door for a new and faster automated method to diagnose sepsis.

Porous liquid crystalline lipid-based nanoparticles are shown here to enable protein analysis in microchip electroseparation by reducing sample adsorption. Additionally, higher stability and reproducibility of the separations were observed. The method was tested by separating green fluorescent protein (GFP) in hot embossed cyclic olefin polymer microchips with integrated fiber grooves for LIF detection. The sample adsorption was indirectly quantified by measuring the height, width and asymmetry of the separation peaks for various concentrations of nanoparticles in the sample and background electrolyte. Without nanoparticles, electropherograms displayed typical signs of extensive adsorption to the channel walls, with low, broad tailing peaks. Higher, narrower more symmetric peaks were generated when 0.5-10% nanoparticles were added, showing a dramatic reduction of sample adsorption. The current through the separation channel decreased with nanoparticle concentration, reducing to half its value when the nanoparticle concentration was increased from 0.5 to 4%. Addition of nanoparticles enabled separations that were otherwise hindered by extensive adsorption, e.g. separation of GFP mutants differing by only one amino acid. It was also observed that increasing the nanoparticle concentration increased the number of impurities that could be resolved in a GFP sample. This indicates that the adsorption is further reduced, and/or that the nanoparticles provide an interacting pseudostationary phase for electro-chromatography.