Instytut Podstawowych Problemów Techniki

Streszczenie:

Herein, we describe a novel method for the assessment of droplet viscosity moving inside microfluidic channels. The method allows for the monitoring of the rate of the continuous growth of bacterial culture. It is based on the analysis of the hydrodynamic resistance of a droplet that is present in a microfluidic channel, which affects its motion. As a result, we were able to observe and quantify the change in the viscosity of the dispersed phase that is caused by the increasing population of interacting bacteria inside a size-limited system. The technique allows for finding the correlation between the viscosity of the medium with a bacterial culture and its optical density. These features, together with the high precision of the measurement, make our viscometer a promising tool for various experiments in the field of analytical chemistry and microbiology, where the rigorous control of the conditions of the reaction and the monitoring of the size of bacterial culture are vital.

Słowa kluczowe:

droplet microfluidics, cell growth, viscosity, rheology, Escherichia coli

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Emerging microfluidic technology has introduced new precision controls over reaction conditions. Owing to the small amount of reagents, microfluidics significantly lowers the cost of carrying a single reaction. Moreover, in two-phase systems, each part of a dispersed fluid can be treated as an independent chemical reactor with a volume from femtoliters to microliters, increasing the throughput. In this work, we propose a microfluidic device that provides continuous recirculation of droplets in a closed loop, maintaining low consumption of oil phase, no cross-contamination, stabilized temperature, a constant condition of gas exchange, dynamic feedback control on droplet volume, and a real-time optical characterization of bacterial growth in a droplet. The channels (tubing) and junction cubes are made of Teflon fluorinated ethylene propylene (FEP) to ensure non-wetting conditions and to prevent the formation of biofilm, which is particularly crucial for biological experiments. We show the design and operation of a novel microfluidic loop with the circular motion of microdroplet reactors monitored with optical sensors and precision temperature controls. We have employed the proposed system for long term monitoring of bacterial growth during the antibiotic chloramphenicol treatment. The proposed system can find applications in a broad field of biomedical diagnostics and therapy.

Słowa kluczowe:

microfluidic loop, bacteria cultures, screening, antibiotic treatment, Escherichia coli

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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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While a significant progress has recently been made in therapy of many cancers, the cure for some high grade cancers, such as the astrocytic cancers, remains elusive. In the latter case, specificity and functionality of the brain tissue limit the options available to surgical and chemotherapeutic treatments. In view of the prospects of reversible blood-brain barrier opening, we have investigated the possibility of a transfection of malignant astrocyte cells with novel graphene oxide nanosheet (GONS) nanocarrier-supported molecular beacons (MB) encoded for the detection of a biomarker survivin (Sur). The behavior of GONS-supported SurMBs (GONS@SurMB) has been characterized using fluorescence spectroscopy, SEM, TEM, Raman spectroscopy, melting transients, resonance elastic light scattering, and cell viability testing. With the GONS@SurMB, we have achieved the limit of detection for tDNA at 37°C: LOD = 24 nM (S/N = 3). In tests with complementary targets and mismatched strands, the proposed fluorescent turn-on GONS@SurMB probes have shown a single-nucleotide polymorphism sensitivity. We have demonstrated the transfection of U-87 MG astrocyte cells with GONS@SurMB nanocarriers which release SurMB upon mRNA detection. The MTT tests indicate that the GONS carrier concentrations up to 133 μg/mL are not cytotoxic to astrocyte cells, although a cell assembly has been encountered at higher carrier concentrations. The GONS alone does not assemble appreciably up to 80 μg/mL. The proposed method can be used for the detection of Sur mRNA in malignant cells and the GONS@SurMB nanocarriers can also be considered as viable candidates for future gene therapy of brain cancers.

Słowa kluczowe:

Survivin detection, Molecular beacon, Graphene oxide nanocarrier, Survivin mRNA, U-87 malignant glioma cells

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Conducting reactions in droplets in microfluidic chips offers several highly attractive characteristics, among others, increased yield and selectivity of chemical syntheses. The use of droplet microfluidic systems in synthetic chemistry is, however, hampered by the intrinsically small throughput of micrometric channels. Here, we verify experimentally the potential to increase throughput via an increase of the scale of the channels.We use the results of these experiments characterizing the processes of (1) generation of droplets, (2) mixing in droplets, (3) inter-phase extraction, and (4) the yield of synthesis of pyrrole, to postulate a number of guidelines for scaling up the throughput of microfluidic droplet systems. In particular, we suggest the rules for maximizing the throughput via an increase of the size of the channels and via parallelization to optimize the throughput of synthesis against the cost of fabrication of the chips and against the kinetic requirements of specific reactions.

Słowa kluczowe:

flow chemistry, microfluidics, synthesis, emulsions, droplets

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Blood is the richest source of diagnostic information. The growing interest in point-of-care analytics prompted several attempts to extract plasma from whole blood in simple diagnostic devices. The simplest method of separation is sedimentation. Here we show the first microfluidic system that uses sedimentation to extract plasma from undiluted blood and integrates execution of liquid assays on the extracted material. We present a microfluidic chip that accepts a small sample (27 μL) of whole blood, separates up to 6 μL of plasma, and uses metered volumes of plasma and of reagent (2-chloro-4-nitrophenyl-α-maltotrioside, CNP-G3) for a liquid enzymatic assay. With a custom designed channel, the system separates blood by sedimentation within few minutes of accepting the sample, mixes it with the reagent, and quantifies spectrophotometrically the product of the enzymatic reaction. As a model demonstration, we show a quantitative enzymatic α-amylase assay that is routinely used in diagnosis of pancreas diseases. The paper reports the design and characterization of the microfluidic device and the results of tests on clinically collected blood samples. The results obtained with the microfluidic system compare well to a reference bench-top analyzer.

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We report automated generation of arbitrary sequences of multiple microdroplets with online and individual control over the number of cores and volumes of all the constituents (cores and shells) of each of the multiple droplets. We show that a given sequence of volumes of the cores always folds to the same final three-dimensional architecture. The method presents the first proof-of-concept for the ability to design the three-dimensional structure of multiple droplets. We discuss the potential use of the technique in the formulation of predetermined distribution of drug release capsules and for automated generation of functional chemical microdroplet networks.

Słowa kluczowe:

multiple droplets, microfluidics

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We present microfluidic modules (traps) that allow us to lock, shift, dose and merge micro-aliquots of liquid precisely. The precision is hard-wired into the geometry of the device: small values of the capillary number guarantee reproducibility of operation over a range of rates of flow that need not be controlled precisely. The modules can be integrated into systems that perform complicated protocols on micro-droplets while not requiring precision in forcing the flow.

Słowa kluczowe:

microfluidics, droplets

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Even at moderate values of Reynolds number [e.g., Re=O(1) ] a curved interface between liquids can induce an abrupt transition between topologically different configurations of laminar flow. Here we show for the first time direct evidence of a sharp transition in the speed of flow of a droplet upon a small increase of the value of the capillary number above a threshold and the associated change of topology of flow. The quantitative results on the dependence of the threshold capillary number on the contrast of viscosities and on the direction of transition cannot be explained by any of the existing theories and call for a new description.

Słowa kluczowe:

speed of droplets, microfluidics, droplets

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Microfluidic droplet-on-demand systems allow the controllable construction of multiple droplets of previously unattainable morphologies. Guided by the diagrams of the possible topologies of double droplets we investigate in detail the vistas to control the morphology of Janus droplets. We also explore and control new morphologies of multiple Janus droplets, i.e., arbitrarily long chains of alternating immiscible segments. Theoretical calculations together with the control offered by the use of automation allow the design of both the topology and the geometry (e.g. curvatures of the interfaces) of the multiple droplets. The ability to rationally design convex–convex, convex–concave and concave–convex segments may be useful in material science, while the ability to tune the distances between the interfaces in the chains of droplets may have applications in designing artificial biochemical signalling networks.

Słowa kluczowe:

multiple droplets, microfluidics

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We report a microfluidic technique for high-throughput generation of droplets of nanolitre volume in parallel channels with online control of the volumes, volume fraction and distribution of droplet volumes with the use of two external valves.

Słowa kluczowe:

microfluidics, droplet generation, droplets

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Streszczenie:

Droplet microfluidic techniques offer an attractive compromise between the throughput (of i.e. reactions per second) and the number of input/output controls needed to control them. Reduction of the number of controls follows from the confinement to essentially one-dimensional flow of slugs in channels which—in turn—relies heavily on the speed of flow of droplets. This speed is a complicated function of numerous parameters, including the volume of droplets (or length L of slugs), their viscosity μd, viscosity μc and rate of flow of the continuous phase, interfacial tension and geometry of the cross-section of the channel. Systematic screens of the impact of these parameters on the speed of droplets remain an open challenge. Here we detail an automated system that screens the speeds of individual droplets at a rate of up to 2000 experiments per hour, with high precision and without human intervention. The results of measurements in channels of square cross-section (of width w = 360 μm) for four different values of the contrast of viscosities λ = μd/μc = 0.3, 1, 3, and 33, wide ranges of values of the capillary number Ca ∈ (10−4, 10−1), and wide ranges of lengths of droplets l = L/w ∈ (0.8, 30) show that the speed of droplets depends significantly both on l and on λ. The dependence on Ca is very strong for λ > 1, while it is less important both for λ ≤ 1 and for λ ≫ 1.

Słowa kluczowe:

microfluidics, capillary flow, droplets

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Streszczenie:

Techniques operating on droplets and treating them as single reactors are very promising due to their advantage: i) lack of dispersion, ii) fast mixing, iii) superior control on kinetics of reactions, iv) easy to paralelization v) small volume The challenge is to control the size and the content of each droplet simultaneously.
We present concept of microfluidic modules that taking advantage of the capillary back pressure allow to trap, dose and realize precise portion of liquid. The precision is encrypted in the geometry of the device and large mismatch of shear stresses and capillary forces. This mismatch guaranties reproducibility over a wide range of rates of flow that need not be controlled precisely. We also demonstrate that these modules allow to built systems that perform complicated protocols precisely while nor precise control.

Słowa kluczowe:

microfluidics, droplets

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