Droplet-based microfluidics

Moragues, Thomas; Arguijo, Diana; Beneyton, Thomas; Modavi, Cyrus; Simutis, Karolis; Abate, Adam R.; deMello, Andrew J.; Densmore, Douglas; Griffiths, Andrew D.

doi:10.1038/s43586-023-00212-3

Cited by 95 publications

(46 citation statements)

References 283 publications

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“…[ 2,32 ] Till now, droplet‐based microfluidic technologies have the capacity for the generation of various droplet forms, including single‐emulsion droplets, double‐emulsion droplets, multiple emulsion droplets, droplets with various biological cargos (e.g., cells, organelles, drugs, and metabolites) and droplets with different structures (e.g., compartmentalization). [ 24a,33 ]…”

Section: Simple Droplets Generated By Microfluidic Platformsmentioning

confidence: 99%

Recent Advances in Microfluidic Technologies for the Construction of Artificial Cells

Tan

Yin

et al. 2023

Adv Funct Materials

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Artificial cells are synthetic constructs that emulate natural cells, with potential applications in areas of energy science, environmental treatment, and the study of life's origins. Nevertheless, the construction of artificial cells is a formidable undertaking, given the intricate nature of natural cells in structures, functions, and working mechanisms. With precise control, high automation, and excellent uniformity, microfluidics has emerged as a promising approach for the construction of artificial cells. This review summarizes the latest microfluidic techniques utilized to construct artificial cells, ranging from simple droplets to sophisticated cell‐inspired systems. These include the generation of droplets, the production of vesicles (lipid‐based and polymer‐based vesicles), the fabrication of polymeric microparticles with various compartments, shapes, and microstructures, as well as the manufacture of sophisticated cell‐inspired systems. The characteristics of different methods for the construction of artificial cells are discussed in detail. Furthermore, the wide‐ranging applications of artificial cells are also showcased. Finally, contemporary obstacles and forthcoming advancements are discussed in the field of microfluidic‐based artificial cells. This review is supposed to stimulate research in the construction of more functional and natural‐like artificial cells, as well as works in the fields of material, biology, environment, medicine, and energy.

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Section: Simple Droplets Generated By Microfluidic Platformsmentioning

confidence: 99%

Recent Advances in Microfluidic Technologies for the Construction of Artificial Cells

Tan

Yin

et al. 2023

Adv Funct Materials

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“…Future developments in biochemical assay technology will need to deliver both miniaturized sample volumes and increased rates of analysis to enable everlarger screening campaigns with ultrahigh throughput. Both of these goals can be fulfilled by encapsulating (bio)chemical assays in water-in-oil microdroplets made in microfluidic devices [1][2] or open systems. 3 Conducting reactions in microdroplets is extremely reagent-saving, with assay volumes in the pico-to femtoliter range.…”

Section: Introductionmentioning

confidence: 99%

“…Droplet-based high-throughput screening methods facilitate analysis of extraordinary sample sizes with throughputs in the tens of kHz range implying a capacity of up to over 100 million independent reactions in a working day. [1][2][4][5][6][7][8][9] This type of screening is advantageous in particular for diagnostic applications, [10][11][12][13] or to identify efficient catalytic systems, 9, 14-15 e.g. by interrogating protein libraries in directed evolution, [16][17][18][19][20][21] and functional metagenomics, [22][23][24] and in strain selections [25][26] or bioprospecting.…”

Section: Introductionmentioning

confidence: 99%

Rapid quantitative assessment of small molecule leakage from microdroplets by flow cytometry and improvement of fluorophore retention in biochemical assays

Zinchenko

Devenish

Hollfelder

2023

Preprint

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Microdroplets are compartments made in the laboratory that allow the miniaturisation of chemical and biological experiments to the femto- to picolitre scale, replacing the classical test tube with a droplet. Ideally containment of the contents of individual droplets would be perfect, but in reality this situation rarely occurs. Instead the leaking of molecules even from intact droplets presents a challenge to the success of miniaturisation and must be assessed on a case-by-case basis. We now present a new method for quantitative determination of leakage: a sheath fluid-free flow cytometer (Guava EasyCyte) is used to directly determine the fluorescence of water-in-oil droplets as a function of time. We validate this method by demonstrating that this assessment of leakage provides a framework for experimental improvements that reduce the leakage of two widely used fluorophores. A 40-fold better retention compared to current protocols is achieved for resorufin with an optimized mix (oil: FC-70, surfactant: 0.1% w/w AZ900C, additive: 1% BSA) to maintain useful retention for up to 130 hours. Likewise leakage of the fluorophore methylumbelliferone is reduced by 75-fold. The availability of a method to quantitate leakage quickly for a variety of experimental conditions will facilitate future applications of droplet-based experiments (e.g. in directed evolution or diagnostics), aid miniaturisation of lab-scale assays into this format, and improve the degrees of freedom in setting up such ultrahigh-throughput experiments.

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“…Herein, we describe a new approach to injection that uses sample introduction by droplet microfluidics to address the cycle time limitations of current autosamplers. , A system was previously developed that coupled an LC-MS with a custom oil-covered droplet array chip using a capillary sampling probe and a 4 nL injection valve, allowing for microfluidic droplets to be sampled and injected in a manner similar to commercially available autosamplers . More recently, an innovative PEEK injector was developed and integrated with a microfluidic chip that allowed individual droplets to be analyzed by chip-LC .…”

Section: Introductionmentioning

confidence: 99%

High-Throughput Liquid Chromatographic Analysis Using a Segmented Flow Injector with a 1 s Cycle Time

Makey,

Diehl,

Xin

et al. 2023

Anal. Chem.

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High-throughput screening (HTS) workflows are revolutionizing many fields, including drug discovery, reaction discovery and optimization, diagnostics, sensing, and enzyme engineering. Liquid chromatography (LC) is commonly deployed during HTS to reduce matrix effects, distinguish isomers, and preconcentrate prior to detection, but LC separation time often limits throughput. Although subsecond LC separations have been demonstrated, they are rarely utilized during HTS due to limitations associated with the speed of common autosamplers. In this work, these limits are overcome by utilizing droplet microfluidics for sample introduction. In the method, a train of samples segmented by air are continuously pumped into the inlet of an LC injection valve that is actuated once each sample fills the sample loop. Coupled with 2.1 mm diameter × 5 mm long columns packed with 2.7 μm superficially porous C18 particles operated at 5 mL/min, the injector enabled separation of 3 components at 1 s/sample and analysis of a 96-well plate in 1.6 min with <2% peak area relative standard deviation. Analyte-dependent carryover was minimized by including wash droplets composed of organic solvent in between sample droplets. High-throughput LC coupled with mass spectrometric detection using the segmented flow injector was applied to a screen of inhibitors of a cytochrome P450-catalyzed hydroxylation reaction. Measurements of the reaction substrate and product concentrations made using fast LC with the segmented flow injector correlated well with measurements made using a more conventional, 3 min LC method. These results demonstrate the potential for droplet microfluidics to be used for sample introduction during high-throughput LC analysis.

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Droplet-based microfluidics

Cited by 95 publications

References 283 publications

Recent Advances in Microfluidic Technologies for the Construction of Artificial Cells

Recent Advances in Microfluidic Technologies for the Construction of Artificial Cells

Rapid quantitative assessment of small molecule leakage from microdroplets by flow cytometry and improvement of fluorophore retention in biochemical assays

High-Throughput Liquid Chromatographic Analysis Using a Segmented Flow Injector with a 1 s Cycle Time

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