Microfluidics has enabled a new era of cellular and molecular assays due to the small length scales, parallelization, and the modularity of various analysis and actuation functions. Droplet microfluidics, in particular, has been instrumental in providing new tools for biology with its ability to quickly and reproducibly generate drops that act as individual reactors. A notable beneficiary of this technology has been single-cell RNA sequencing, which has revealed new heterogeneities and interactions for the fundamental unit of life. However, viruses far surpass the diversity of cellular life, affect the dynamics of all ecosystems, and are a chronic source of global health crises. Despite their impact on the world, high-throughput and high-resolution viral profiling has been difficult, with conventional methods being limited to population-level averaging, large sample volumes, and few cultivable hosts. Consequently, most viruses have not been identified and studied. Droplet microfluidics holds the potential to address many of these limitations and offers new levels of sensitivity and throughput for virology. This Feature highlights recent efforts that have applied droplet microfluidics to the detection and study of viruses, including for diagnostics, virus-host interactions, and cell-independent virus assays. In combination with traditional virology methods, droplet microfluidics should prove a potent tool toward achieving a better understanding of the most abundant biological species on Earth.
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http://dx.doi.org/10.1021/acs.analchem.2c00615 | DOI Listing |
Anal Chem
January 2025
Mechanical Engineering, Faculty of Engineering and Physical Sciences, University of Southampton, Southampton SO17 1BJ, U.K.
Total free thiols are an important marker of the whole-body redox state, which has been shown to be associated with clinical outcome in health and disease. Recent investigations have suggested that increased insight may be gained by monitoring alterations of redox state in response to exercise and hypoxia and to monitor redox trajectories in disease settings. However, conducting such studies is challenging due to the requirement for repeated venous blood sampling and intensive lab work.
View Article and Find Full Text PDFPNAS Nexus
January 2025
Institute of Bioengineering, School of Engineering, École Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, Switzerland.
Here, we present Link-Seq, a highly efficient droplet microfluidic method for combined sequencing of antibody-encoding genes and the transcriptome of individual B cells at large scale. The method is based on 3' barcoding of the transcriptome and subsequent single-molecule PCR in droplets, which freely shift the barcode along specific gene regions, such as the antibody heavy- and light-chain genes. Using the immune repertoire of COVID-19 patients and healthy donors as a model system, we obtain up to 91.
View Article and Find Full Text PDFHeliyon
January 2025
Institute for Nanomaterials, Advanced Technologies and Innovation, Technical University of Liberec, 46117, Liberec, Czech Republic.
Droplet coalescence in microchannels is a complex phenomenon influenced by various parameters such as droplet size, velocity, liquid surface tension, and droplet-droplet spacing. In this study, we thoroughly investigate the impact of these control parameters on droplet coalescence dynamics within a sudden expansion microchannel using two distinct numerical methods. Initially, we employ the boundary element method to solve the Brinkman integral equation, providing detailed insights into the underlying physics of droplet coalescence.
View Article and Find Full Text PDFSmall
January 2025
School of Biomedical Sciences, LKS Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Fluorescent light-up aptamer/fluorogen pairs are powerful tools for tracking RNA in the cell, however limitations in thermostability and fluorescence intensity exist. Current in vitro selection techniques struggle to mimic complex intracellular environments, limiting in vivo biomolecule functionality. Taking inspiration from microenvironment-dependent RNA folding observed in cells and organelle-mimicking droplets, an efficient system is created that uses microscale heated water droplets to simulate intracellular conditions, effectively replicating the intracellular RNA folding landscape.
View Article and Find Full Text PDFMicrosyst Nanoeng
January 2025
Guangdong Provincial Key Laboratory of Sensor Technology and Biomedical Instrument, School of Biomedical Engineering, Shenzhen Campus of Sun Yat-Sen University, 518000, Shenzhen, China.
Advancements in screening technologies employing small organisms have enabled deep profiling of compounds in vivo. However, current strategies for phenotyping of behaving animals, such as zebrafish, typically involve tedious manipulations. Here, we develop and validate a fully automated in vivo screening system (AISS) that integrates microfluidic technology and computer-vision-based control methods to enable rapid evaluation of biological responses of non-anesthetized zebrafish to molecular gradients.
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