Droplet-based microfluidics

Summary

Droplet-based microfluidics manipulate discrete volumes of fluids in immiscible phases with low Reynolds number and laminar flow regimes. Interest in droplet-based microfluidics systems has been growing substantially in past decades. Microdroplets offer the feasibility of handling miniature volumes (μl to fl) of fluids conveniently, provide better mixing, encapsulation, sorting, sensing and are suitable for high throughput experiments. Two immiscible phases used for the droplet based systems are referred to as the continuous phase (medium in which droplets flow) and dispersed phase (the droplet phase). Droplet formation methods In order for droplet formation to occur, two immiscible phases, referred to as the continuous phase (medium in which droplets are generated) and dispersed phase (the droplet phase), must be used. The size of the generated droplets is mainly controlled by the flow rate ratio of the continuous phas

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https://en.wikipedia.org/wiki/Droplet-based_microfluidics

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A microfluidic Braille valve platform for on-demand production, combinatorial screening and sorting of chemically distinct droplets

Wenwei Ma, Christoph Merten, Vida Vafaizadeh

Droplet microfluidics is a powerful tool for a variety of biological applications including single-cell genetics, antibody discovery and directed evolution. All these applications make use of genetic libraries, illustrating the difficulty of generating chemically distinct droplets for screening applications. This protocol describes our Braille Display valving platform for on-demand generation of droplets with different chemical contents (16 different reagents and combinations thereof), as well as sorting droplets with different chemical properties, on the basis of fluorescence signals. The Braille Display platform is compact, versatile and cost efficient (only similar to US$1,000 on top of a standard droplet microfluidics setup). The procedure includes manufacturing of microfluidic chips, assembly of custom hardware, co-encapsulation of cells and drugs into droplets, fluorescence detection of readout signals and data analysis using shared, freely available LabVIEW and Python packages. As a first application, we demonstrate the complete workflow for screening cancer cell drug sensitivities toward 74 conditions. Furthermore, we describe here an assay enabling the normalization of the observed drug sensitivity to the number of cancer cells per droplet, which additionally increases the robustness of the system. As a second application, we also demonstrate the sorting of droplets according to enzymatic activity. The drug screening application can be completed within 2 d; droplet sorting takes similar to 1 d; and all preparatory steps for manufacturing molds, chips and setting up the Braille controller can be accomplished within 1 week.

NATURE PORTFOLIO2022

A new Additive-Manufacturing (AM) or 3D printing concept is proposed to improve the printing resolution for metal additive manufacturing in the frame of the SFA-AM project, Powder Focusing for Beam-Induced Laser 3D Printing. The project aims to transport small powder particles (largest diameter < 10 um) at a high throughput to a spot smaller than 20 µm. To realize the objective, a metal ion included liquid is suggested as a source medium similar to the inkjet process, and the droplet is employed as a container for the metal ion transport. The key challenges of achieving this goal are 1) droplet size control and 2) efficient microwave heating that can dry the liquid portion of the dynamic droplet at a high-speed (3 m/s). The most demanding part is the development of a miniaturized, very concentrated field density microwave resonator where the droplet is passing through. This follows from the extremely short interaction time between the microwave and the ejected droplets (research: 0.5 ms) which results from the requirement for a small system size for its integration. The primary medium examined for the experiment is water due to 1) a high dielectric loss proportional to the drying efficiency and 2) avoiding explosion risks at the laser sintering stage compared to organic solutions. Various droplet generation conditions were investigated to determine the optimum conditions for the custom droplet generator, in particular, two key parameters were focused on: frequency and flow rate. The droplets obtained were analyzed with respect to size, gap distance, and linearity affecting the uniformity of the AM process. Moreover, Navier-Stokes-based fluid dynamics simulations enabled understanding of droplets behaviors, especially break-up formation. Eventually, the optimal condition for producing the smallest droplets with a diameter of 100 µm is determined to be 15 kHz and 0.5 ml/min. Such droplets loaded with metallic particles could result in smaller than 20 um of remaining metal powders aggregate after the drying process.A new TEM (Transverse ElectroMagnetic) resonator has been developed for drying and sensing applications. A full-wave electromagnetic simulation software, CST Microwave Studio, supported the working mode of the device. Firstly, it is used for the dielectric characterizations of different water-ethanol mixtures contained in a micro capillary. The sensing relies on electric field perturbation due to the inserted sample, and is measured based on the change of both resonance frequency and Q-factor. These sample-dependent variations were analyzed to estimate the complex permittivity using different methods such as the Perturbation Method (PM), Least-Square Model (LSM), and Log-Linear Model (LLM). This work demonstrates that the proposed microwave module is capable of sensing nano-liter volume ranges even though the samples only partially influence the sensing area (error < 13 % in permittivity). In microwave heating, the developed resonator achieved a temperature increase of 28 K in the microwave exposal time of 0.5 ms when the microwave power of 45 W was applied. Finally, numerical models have been proposed to estimate the maximum temperature of the initial droplet before cooling starts attributable to heat and mass transfer. This model well corresponds to the microwave electromagnetic simulation resulting in similar temperature deviations. It helps to determine the net heating efficiency of the microwave device.

EPFL2022

The invention provides a method for quantifying the number of target entities that were successfully encapsulated into a microfluidic compartment such as a microfluidic droplet. The invention is predicated upon a continuous quantifiable detectable signal corresponding to the quantity of entities encapsulated in a microfluidic compartment. The present invention is useful in the context of high throughput microfluidic screening approaches which require thorough signal normalization in order to reliably improve signal to noise ratio and to reliably identify screening hits.

2022

EPFL2022