Formation of bubbles and droplets in microfluidic systems

This mini-review reports the recent advances in the hydrodynamic techniques for formation of bubbles of gas in liquid in microfluidic systems. Systems comprising ducts that have widths of the order of 100 micrometers produce suspensions of bubbles with narrow size distributions. Certain of these systems have the ability to tune the volume fraction of the gaseous phase – over the whole range from zero to one. The rate of flow of the liquids through the devices determines the mechanism of formation of the bubbles – from break-up controlled by the rate of flow of the liquid (at low capillary numbers, and in the presence of strong confinement by the walls of the microchannels), to dynamics dominated by inertial effects (at high Weber numbers). The region of transition between these two regimes exhibits nonlinear behaviours, with period doubling cascades and irregular bubbling as prominent examples. Microfluidic systems provide new and uniquely controlled methods for generation of bubbles, and offer potential applications in micro-flow chemical processing, synthesis of materials, and fluidic optics.The U.S. Department of Energy DE-FG02- 00ER45852Foundation for Polish ScienceMinisterio de Educación y Ciencia de España DPI2002-04305-C02-02

Fast selective trapping and release of picoliter droplets in a 3D microfluidic PDMS multi-trap system with bubbles

The selective manipulation and incubation of individual picoliter drops in high-throughput droplet based microfluidic devices still remains challenging. We used a surface acoustic wave (SAW) to induce a bubble in a 3D designed multi-trap polydimethylsiloxane (PDMS) device to manipulate multiple droplets and demonstrate the selection, incubation and on-demand release of aqueous droplets from a continuous oil flow. By controlling the position of the acoustic actuation, individual droplets are addressed and selectively released from a droplet stream of 460 drops per s. A complete trapping and releasing cycle can be as short as 70 ms and has no upper limit for incubation time. We characterize the fluidic function of the hybrid device in terms of electric power, pulse duration and acoustic path

Tuna-step: Tunable Parallelized Step Emulsification for the Generation of Droplets with Dynamic Volume Control to 3D Print Functionally Graded Porous Materials

We present Tuna-step, a novel microfluidic module based on step emulsification that allows for reliable generation of droplets of different sizes. Until now, sizes of droplets generated with step emulsification were hard-wired into the geometry of the step emulsification nozzle. To overcome this, we incorporate a thin membrane underneath the step nozzle that can be actuated by pressure, enabling the tuning of the nozzle size on-demand. By controllably reducing the height of the nozzle, we successfully achieved a three-order-of-magnitude variation in droplet volume without any adjustment of the flow rates of the two phases. We further enhanced our system by developing and applying a new hydrophilic surface modification, ensuring long-term stability and preventing swelling of the device when generating Oil-in-Water droplets. We used our system in the manufacturing of functionally graded soft materials with adjustable porosity and material content. By combining our microfluidic device with a custom 3D printer, we generated and extruded Oil-in-Water emulsions in an agarose gel bath, allowing for the creation of unique self-standing 3D hydrogel structures with porosity decoupled from flow rate and with composition gradients of external phases. We upscaled Tuna-step by setting 14 actuatable nozzles in parallel, offering a step-emulsification-based single chip solution that can accommodate various requirements in terms of throughput, droplet volumes, flow rates, and surface chemistry

Droplet-based digital antibiotic susceptibility screen reveals single-cell clonal heteroresistance in an isogenic bacterial population

Since antibiotic resistance is a major threat to global health, recent observations that the traditional test of minimum inhibitory concentration (MIC) is not informative enough to guide effective antibiotic treatment are alarming. Bacterial heteroresistance, in which seemingly susceptible isogenic bacterial populations contain resistant sub-populations, underlies much of this challenge. To close this gap, here we developed a droplet-based digital MIC screen that constitutes a practical analytical platform for quantifying the single-cell distribution of phenotypic responses to antibiotics, as well as for measuring inoculum effect with high accuracy. We found that antibiotic efficacy is determined by the amount of antibiotic used per bacterial colony forming unit (CFU), not by the absolute antibiotic concentration, as shown by the treatment of beta-lactamase-carrying Escherichia coli with cefotaxime. We also noted that cells exhibited a pronounced clustering phenotype when exposed to near-inhibitory amounts of cefotaxime. Overall, our method facilitates research into the interplay between heteroresistance and antibiotic efficacy, as well as research into the origin and stimulation of heterogeneity by exposure to antibiotics. Due to the absolute bacteria quantification in this digital assay, our method provides a platform for developing reference MIC assays that are robust against inoculum-density variations

High-Throughput Monitoring of Bacterial Cell Density in Nanoliter Droplets: Label-Free Detection of Unmodified Gram-Positive and Gram-Negative Bacteria

Droplet microfluidics disrupted analytical biology with the introduction of digital polymerase chain reaction and single-cell sequencing. The same technology may also bring important innovation in the analysis of bacteria, including antibiotic susceptibility testing at the single-cell level. Still, despite promising demonstrations, the lack of a high-throughput label-free method of detecting bacteria in nanoliter droplets prohibits analysis of the most interesting strains and widespread use of droplet technologies in analytical microbiology. We use a sensitive and fast measurement of scattered light from nanoliter droplets to demonstrate reliable detection of the proliferation of encapsulated bacteria. We verify the sensitivity of the method by simultaneous readout of fluorescent signals from bacteria expressing fluorescent proteins and demonstrate label-free readout on unlabeled Gram-negative and Gram-positive species. Our approach requires neither genetic modification of the cells nor the addition of chemical markers of metabolism. It is compatible with a wide range of bacterial species of clinical, research, and industrial interest, opening the microfluidic droplet technologies for adaptation in these fields

Highly ordered and tunable polyHIPEs by using microfluidics

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