Instytut Podstawowych Problemów Techniki

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Two decades of research on droplet formation in microchannels have led to the widely accepted view that droplets form through the squeezing mechanism when interfacial forces dominate over viscous forces. The initially surprising finding that the volume of the droplets is insensitive to the relative importance of these two forces is nowadays well understood from the constrained deformation of the droplet interface during formation. In this work, we show a lower limit of the squeezing mechanism for droplets produced in microfluidic cross-junctions. Below this limit, in the leaking regime, which was recently discovered for droplets produced in T-junctions, the volume of the produced droplets strongly depends on the relative importance of interfacial and viscous forces, as captured by the capillary number. We reveal a fundamental difference in the mechanisms at play in the leaking regime between T- and cross-junctions. In cross-junctions, the droplet neck elongates substantially, and unlike the case of the T-junction, the magnitude of this elongation depends strongly on the value of the capillary number. This elongation significantly affects the final droplet volume in a low capillary number regime. Generalizing the classical squeezing law by lifting the original assumptions and incorporating both identified mechanisms of leaking through gutters and neck elongation, we derive a model for droplet formation and show that it agrees with our experiments.

Słowa kluczowe:

Microfluidics,Cross-junction,Flow-focusing device,Droplet formation,Two-phase flow,Scaling law,Squeezing regime

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We present a novel microfluidic system that produces the required concentration of a reagent in a single droplet or that produces a sequence of droplets with a defined periodic distribution of concentrations. We use digital algorithms that, through a series of simple operations, such as merging and splitting droplets, ensure superior precision, repeatability and flexibility in concentration setting. Unlike Digital microfluidic (DMF) systems based on electrowetting on dielectric (EWOD) commonly used to implement digital algorithms in the droplet world, our approach is based on much more available channel-based microfluidics operated by programmable syringe pumps. Furthermore, the small footprint of our system makes it easy to integrate with other structures of microfluidic networks. Thus, this technique is a comprehensive component that can be built into the microfluidic networks executing laboratory analytical tasks in chemistry or biology to enrich their performance and offer new functionalities.

Słowa kluczowe:

droplet-based microfluidics, microfluidic traps, droplet manipulation, concentration tuning, concentration gradient

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Stored red blood cells (RBCs) incur biochemical and morphological changes, collectively termed the storage lesion. Functionally, the storage lesion manifests as slower oxygen unloading from RBCs, which may compromise the efficacy of transfusions where the clinical imperative is to rapidly boost oxygen delivery to tissues. Recent analysis of large real-world data linked longer storage with increased recipient mortality. Biochemical rejuvenation with a formulation of adenosine, inosine, and pyruvate can restore gas-handling properties, but its implementation is impractical for most clinical scenarios. We tested whether storage under hypoxia, previously shown to slow biochemical degradation, also preserves gas-handling properties of RBCs. A microfluidic chamber, designed to rapidly switch between oxygenated and anoxic superfusates, was used for single-cell oxygen saturation imaging on samples stored for up to 49 days. Aliquots were also analyzed flow cytometrically for side-scatter (a proposed proxy of O2 unloading kinetics), metabolomics, lipidomics, and redox proteomics. For benchmarking, units were biochemically rejuvenated at 4 weeks of standard storage. Hypoxic storage hastened O2 unloading in units stored to 35 days, an effect that correlated with side-scatter but was not linked to posttranslational modifications of hemoglobin. Although hypoxic storage and rejuvenation produced distinct biochemical changes, a subset of metabolites including pyruvate, sedoheptulose 1-phosphate, and 2/3 phospho-d-glycerate, was a common signature that correlated with changes in O2 unloading. Correlations between gas handling and lipidomic changes were modest. Thus, hypoxic storage of RBCs preserves key metabolic pathways and O2 exchange properties, thereby improving the functional quality of blood products and potentially influencing transfusion outcomes.

Słowa kluczowe:

hypoxia, Hemanext, erythrocyte, hemoglobin, oxidative stress, microfluidics

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Epithelial folding is a fundamental process where initially flat monolayers transform into functional 3D structures. This protocol details fabrication steps for a polycarbonate microfluidic platform which enables triggering epithelial folds that recapitulate stereotypical cell shape changes and folding-associated mechanical stresses. We describe the steps for cell seeding to form a monolayer on the chip, and subsequent approach to trigger calcium waves in the epithelial monolayer through local epithelial deformation. Lastly, we outline quantitative analysis steps of the epithelial response.

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Cell shape dynamics during development is tightly regulated and coordinated with cell fate determination. Triggered by an interplay between biochemical and mechanical signals, epithelia form complex tissues by undergoing coordinated cell shape changes, but how such spatiotemporal coordination is controlled remains an open question. To dissect biochemical signaling from purely mechanical cues, we developed a microfluidic system that experimentally triggers epithelial folding to recapitulate stereotypic deformations observed in vivo. Using this system, we observe that the apical or basal direction of folding results in strikingly different mechanical states at the fold boundary, where the balance between tissue tension and torque (arising from the imposed curvature) controls the spread of folding-induced calcium waves at a short timescale and induces spatial patterns of gene expression at longer timescales. Our work uncovers that folding-associated gradients of cell shape and their resulting mechanical stresses direct spatially distinct biochemical responses within the monolayer.

Słowa kluczowe:

epithelial morphogenesis, epithelial folding, tension, calcium waves, microfluidics, RNAseq

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Here, we show the successful implementation of advanced sequential logic in droplet microfluidics, whose principles rely on capillary wells establishing stationary states, where droplets can communicate remotely via pressure impulses, influencing each other and switching the device states. All logic operations perform spontaneously due to the utilization of nothing more than capillary–hydrodynamic interactions, inherent for the confined biphasic flow. Our approach offers integration feasibility allowing to encode unprecedentedly long algorithms, e.g., 1000-droplet counting. This work has the potential for the advancement of liquid computers and thereby could participate in the development of the next generation of portable microfluidic systems with embedded control, enabling applications from single-cell analysis and biochemical assays to materials science.

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In the present paper, we provide evidence of the vital impact of inertia on the flow in microfluidic networks, which is disclosed by the appearance of nonlinear velocity–pressure coupling. The experiments and numerical analysis of microfluidic junctions within the range of moderate Reynolds number (1 < Re < 250) revealed that inertial effects are of high relevance when Re > 10. Thus, our results estimate the applicability limit of the linear relationship between the flow rate and pressure drop in channels, commonly described by the so-called hydraulic resistance. Herein, we show that neglecting the nonlinear in their nature inertial effects can make such linear resistance-based approximation mistaken for the network operating beyond Re < 10. In the course of our research, we investigated the distribution of flows in connections of three channels in two flow modes. In the splitting mode, the flow from a common channel divides between two outputs, while in the merging mode, streams from two channels join together in a common duct. We tested a wide range of junction geometries characterized by parameters such as: (1) the angle between bifurcating channels (45°, 90°, 135° and 180°); (2) angle of the common channel relative to bifurcating channels (varied within the available range); (3) ratio of lengths of bifurcating channels (up to 8). The research revealed that the inertial effects strongly depend on angles between the channels. Additionally, we observed substantial differences between the distributions of flows in the splitting and merging modes in the same geometries, which reflects the non-reversibility of the motion of an inertial fluid. The promising aspect of our research is that for some combinations of both lengths and angles of the channels, the inertial contributions balance each other in such a way that the equations recover their linear character. In such an optimal configuration, the dependence on Reynolds number can be effectively mitigated.

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While shear emulsification is a well understood industrial process, geometrical confinement in microfluidic systems introduces fascinating complexity, so far prohibiting complete understanding of droplet formation. The size of confined droplets is controlled by the ratio between shear and capillary forces when both are of the same order, in a regime known as jetting, while being surprisingly insensitive to this ratio when shear is orders of magnitude smaller than capillary forces, in a regime known as squeezing. Here, we reveal that further reduction of—already negligibly small—shear unexpectedly re-introduces the dependence of droplet size on shear/capillary-force ratio. For the first time we formally account for the flow around forming droplets, to predict and discover experimentally an additional regime—leaking. Our model predicts droplet size and characterizes the transitions from leaking into squeezing and from squeezing into jetting, unifying the description for confined droplet generation, and offering a practical guide for applications.

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We present a novel type of microfluidic bifurcating junctions which fixes the droplet's route. Unlike in regular junctions, where a droplet chooses one of two outputs depending on the (often instantaneous) flow distribution, our modifications direct droplets only to one preferred outlet. As we show, this solution works properly regardless of the variations of flow distribution in a wide range of its amplitude. Such modified junctions allow for the encoding of the droplet's traffic in the geometry of the device. We compare in a series of experiments different junctions having channels of uniform square cross section. Our observations revealed that a small, local modification of the junction in the form of an additional shallow slit imposes a significant consequence for the flow of droplets at an entire microfluidic network's scale. Another interesting and helpful feature of these new junctions is that they keep the integrity of long droplets, unlike regular junctions, which tend to split long droplets. Our experimental investigations revealed a complex transformation of the long droplet during its transfer through the modified junction. We show that this transformation resembles the Baker's transform and can be used for the enhancement of mixing inside the droplets. Finally, we show two examples of microfluidic devices where the deterministic character of these modified junctions is utilized to obtain new, non-trivial functionalities. This approach can be used for the engineering of microfluidic devices with embedded procedures replacing active elements like valves or magnetic/electric fields.

Słowa kluczowe:

droplet, microfluidics, two-phase, manipulations

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Multiple pipetting is a standard laboratory procedure resulting in the compartmentalisation of a liquid sample. Microfluidics offers techniques which can replace this process by the use of tiny droplets. Passive manipulation on droplets is an interesting and promising approach for the design of microfluidic devices which on one hand are easy-to-use and on the other, execute complex laboratory procedures. We present a comprehensive study of the geometry of microfluidic components which encode different operations on droplets into the structure of the device. The understanding of hydrodynamic interactions between the continuous flow and a droplet travelling through confined space of nontrivial microfluidic geometries is crucial for a rational and efficient design of new generation of modular microfluidic processors with embedded instructions.

Słowa kluczowe:

microfluidics, two-phase flows, droplets

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