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High-throughput single-cell activity-based screening and sequencing of antibodies using droplet microfluidics
Annabelle Gérard, Adam Woolfe, […]Colin Brenan
Nat Biotechnol. - 38 715–721 - https://doi.org/10.1038/s41587-020-0466-7 - 2020
Mining the antibody repertoire of plasma cells and plasmablasts could enable the discovery of useful antibodies for therapeutic or research purposes1. We present a method for high-throughput, single-cell screening of IgG-secreting primary cells to characterize antibody binding to soluble and membrane-bound antigens. CelliGO is a droplet microfluidics system that combines high-throughput screening for IgG activity, using fluorescence-based in-droplet single-cell bioassays2, with sequencing of paired antibody V genes, using in-droplet single-cell barcoded reverse transcription. We analyzed IgG repertoire diversity, clonal expansion and somatic hypermutation in cells from mice immunized with a vaccine target, a multifunctional enzyme or a membrane-bound cancer target. Immunization with these antigens yielded 100–1,000 IgG sequences per mouse. We generated 77 recombinant antibodies from the identified sequences and found that 93% recognized the soluble antigen and 14% the membrane antigen. The platform also allowed recovery of ~450–900 IgG sequences from ~2,200 IgG-secreting activated human memory B cells, activated ex vivo, demonstrating its versatility.
Metabolic cost of rapid adaptation of single yeast cells
Gabrielle Woronoff, Philippe Nghe, Jean Baudry, Laurent Boitard, Erez Braun, Andrew D. Griffiths, and Jérôme Bibette
PNAS - 117 (20) 10660-10666 - https://doi.org/10.1073/pnas.1913767117 - 2020
Cells can rapidly adapt to changing environments through nongenetic processes; however, the metabolic cost of such adaptation has never been considered. Here we demonstrate metabolic coupling in a remarkable, rapid adaptation process (1 in 1,000 cells adapt per hour) by simultaneously measuring metabolism and division of thousands of individual Saccharomyces cerevisiae cells using a droplet microfluidic system: droplets containing single cells are immobilized in a two-dimensional (2D) array, with osmotically induced changes in droplet volume being used to measure cell metabolism, while simultaneously imaging the cells to measure division. Following a severe challenge, most cells, while not dividing, continue to metabolize, displaying a remarkably wide diversity of metabolic trajectories from which adaptation events can be anticipated. Adaptation requires a characteristic amount of energy, indicating that it is an active process. The demonstration that metabolic trajectories predict a priori adaptation events provides evidence of tight energetic coupling between metabolism and regulatory reorganization in adaptation. This process allows S. cerevisiae to adapt on a physiological timescale, but related phenomena may also be important in other processes, such as cellular differentiation, cellular reprogramming, and the emergence of drug resistance in cancer.
Microfluidics Mediated Production of Foams for Biomedical Applications
Ilham Maimouni Cesare M. Cejas ,Janine Cossy ,Patrick Tabeling and Maria Russo
Review - 11(1) 83 - doi.org/10.3390/mi11010083 - 2020
Within the last decade, there has been increasing interest in liquid and solid foams for several industrial uses. In the biomedical field, liquid foams can be used as delivery systems for dermatological treatments, for example, whereas solid foams are frequently used as scaffolds for tissue engineering and drug screening. Most of the foam functionalities are largely correlated to their mechanical properties and their structure, especially bubble/pore size, shape, and interconnectivity. However, the majority of conventional foaming fabrication techniques lack pore size control which can induce important inhomogeneities in the foams and subsequently decrease their performance. In this perspective, new advanced technologies have been introduced, such as microfluidics, which offers a highly controlled production, allowing for design customization of both liquid foams and solid foams obtained through liquid-templating. This short review explores both the fabrication and the characterization of foams, with a focus on solid polymer foams, and sheds the light on how microfluidics can overcome some existing limitations, playing a crucial role in their production for biomedical applications, especially as scaffolds in tissue engineering. View Full-Text
Microfluidics Mediated Production of Foams for Biomedical Applications
Maria Russo, Zacharias Amara, Johan Fenneteau, Pauline Chaumont-Olive, Ilham Maimouni, Patrick Tabeling and Janine Cossy
Review - 56 5807-5810 - doi.org/10.1039/D0CC02182B - 2020
Liquid foams exhibiting long-term stability are a key-challenge in material design. Based on this perspective, new pyridinium polyfluorinated surfactants were synthesized from simple building blocks enabling unusually stable liquid foams. While the batch-generated foams were used for qualitative foaming evaluation, microfluidics allowed a quantitative insight into the aging effects of monodisperse foams.
Integration of a soft dielectric composite into a cantilever beam for mechanical energy harvesting, comparison between capacitive and triboelectric transducers
Mickaël Pruvost, Wilbert J. Smit, Cécile Monteux, Pablo Del Corro, Isabelle Dufour, Cédric Ayela, Philippe Poulin & Annie Colin
Scientific Reports - 10 20681 - - 2020
Flexible dielectrics that harvest mechanical energy via electrostatic effects are excellent candidates as power sources for wearable electronics or autonomous sensors. The integration of a soft dielectric composite (polydimethylsiloxane PDMS-carbon black CB) into two mechanical energy harvesters is here presented. Both are based on a similar cantilever beam but work on different harvesting principles: variable capacitor and triboelectricity. We show that without an external bias the triboelectric beam harvests a net density power of 0.3 μW/cm2 under a sinusoidal acceleration of 3.9g at 40 Hz. In a variable capacitor configuration, a bias of 0.15 V/μm is required to get the same energy harvesting performance under the same working conditions. As variable capacitors’ harvesting performance are quadratically dependent on the applied bias, increasing the bias allows the system to harvest energy much more efficiently than the triboelectric one. The present results make CB/PDMS composites promising for autonomous portable multifunctional systems and intelligent sensors.
Emulsion Destabilization by Squeeze Flow
Riande I Dekker, Antoine Deblais, Krassimir P Velikov , Peter Veenstra , Annie Colin , Hamid Kellay , Willem K Kegel , Daniel Bonn
Langmuir - 36(27) 7795-7800 - doi: 10.1021/acs.langmuir.0c00759 - 2020
There is a large debate on the destabilization mechanism of emulsions. We present a simple technique using mechanical compression to destabilize oil-in-water emulsions. Upon compression of the emulsion, the continuous aqueous phase is squeezed out, while the dispersed oil phase progressively deforms from circular to honeycomb-like shapes. The films that separate the oil droplets are observed to thin and break at a critical oil/water ratio, leading to coalescence events. Electrostatic interactions and local droplet rearrangements do not determine film rupture. Instead, the destabilization occurs like an avalanche propagating through the system, starting at areas where the film thickness is smallest.
Emulsion Destabilization by Squeeze Flow
Riande I Dekker, Antoine Deblais, Krassimir P Velikov , Peter Veenstra , Annie Colin , Hamid Kellay , Willem K Kegel , Daniel Bonn
Langmuir - 36(27) 7795-7800 - doi: 10.1021/acs.langmuir.0c00759 - 2020
There is a large debate on the destabilization mechanism of emulsions. We present a simple technique using mechanical compression to destabilize oil-in-water emulsions. Upon compression of the emulsion, the continuous aqueous phase is squeezed out, while the dispersed oil phase progressively deforms from circular to honeycomb-like shapes. The films that separate the oil droplets are observed to thin and break at a critical oil/water ratio, leading to coalescence events. Electrostatic interactions and local droplet rearrangements do not determine film rupture. Instead, the destabilization occurs like an avalanche propagating through the system, starting at areas where the film thickness is smallest.
Emulsion Destabilization by Squeeze Flow
Riande I Dekker, Antoine Deblais, Krassimir P Velikov , Peter Veenstra , Annie Colin , Hamid Kellay , Willem K Kegel , Daniel Bonn
Langmuir - 36(27) 7795-7800 - doi: 10.1021/acs.langmuir.0c00759 - 2020
There is a large debate on the destabilization mechanism of emulsions. We present a simple technique using mechanical compression to destabilize oil-in-water emulsions. Upon compression of the emulsion, the continuous aqueous phase is squeezed out, while the dispersed oil phase progressively deforms from circular to honeycomb-like shapes. The films that separate the oil droplets are observed to thin and break at a critical oil/water ratio, leading to coalescence events. Electrostatic interactions and local droplet rearrangements do not determine film rupture. Instead, the destabilization occurs like an avalanche propagating through the system, starting at areas where the film thickness is smallest.
Density waves in shear-thickening suspensions
Guillaume Ovarlez, Anh Vu Nguyen Le2, Wilbert J. Smit2, Abdoulaye Fall
Science Advances - 6 16 - DOI: 10.1126/sciadv.aay5589 - 2020
Shear thickening corresponds to an increase of the viscosity as a function of the shear rate. It is observed in many concentrated suspensions in nature and industry: water or oil saturated sediments, crystal-bearing magma, fresh concrete, silica suspensions, and cornstarch mixtures. Here, we reveal how shear-thickening suspensions flow, shedding light onto as yet non-understood complex dynamics reported in the literature. When shear thickening is important, we show the existence of density fluctuations that appear as periodic waves moving in the direction of flow and breaking azimuthal symmetry. They come with strong normal stress fluctuations of the same periodicity. The flow includes small areas of normal stresses of the order of tens of kilopascals and areas of normal stresses of the order of hundreds of pascals. These stress inhomogeneities could play an important role in the damage caused by thickening fluids in the industry.
Shear thickening in dense non-Brownian suspensions: Viscous to inertial transition
Y. Madraki, A. Oakley, A. Nguyen Le, A. Colin, G. Ovarlez, and S. Hormozi
Journal of Rheology - 64 27 - doi.org/10.1122/1.5129680 - 2020
We present an experimental study on the viscous to inertial mode of shear thickening in dense non-Brownian suspensions. We design a model suspension consisting of monosized spherical particles within a Newtonian suspending fluid. We develop a protocol for the rheological characterization of dense suspensions using the conventional rheometry technique. Our results provide constitutive laws for suspensions with solid volume fractions close to jamming when both viscous and inertial effects at the particle scale are present. We perform atomic force microscopy to measure forces between the particles immersed in the suspending fluid and show that our system of study corresponds to the frictionless regime of dense suspensions in which viscous and collisional forces dissipate the energy. Finally, we show that the proposed empirical constitutive laws, when approaching jamming, predict the dynamics of dense suspensions in a transient boundary driven flow.

581 publications.