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In the past five years, droplet microfluidic techniques have unlocked new opportunities for the high-throughput genome-wide analysis of single cells, transforming our understanding of cellular diversity and function. However, the field lacks an accessible method to screen and sort droplets based on cellular phenotype upstream of genetic analysis, particularly for large and complex cells. To meet this need, we developed Dropception, a robust, easy-to-use workflow for precise single-cell encapsulation into picoliter-scale double emulsion droplets compatible with high-throughput screening via fluorescence-activated cell sorting (FACS). We demonstrate the capabilities of this method by encapsulating five standardized mammalian cell lines of varying sizes and morphologies as well as a heterogeneous cell mixture of a whole dissociated flatworm (5-25 μm in diameter) within highly monodisperse double emulsions (35 μm in diameter). We optimize for preferential encapsulation of single cells with extremely low multiple-cell loading events ( less then 2% of cell-containing droplets), thereby allowing direct linkage of cellular phenotype to genotype. Across all cell lines, cell loading efficiency approaches the theoretical limit with no observable bias by cell size. FACS measurements reveal the ability to discriminate empty droplets from those containing cells with good agreement to single-cell occupancies quantified via microscopy, establishing robust droplet screening at single-cell resolution. High-throughput FACS screening of cellular picoreactors has the potential to shift the landscape of single-cell droplet microfluidics by expanding the repertoire of current nucleic acid droplet assays to include functional phenotyping.Growing antimicrobial resistance (AMR) is a serious global threat to human health. Current methods to detect resistance include phenotypic antibiotic sensitivity testing (AST), which measures bacterial growth and is therefore hampered by a slow time to obtain results (∼12-24 h). Therefore, new rapid phenotypic methods for AST are urgently needed. Nanomechanical cantilever sensors have recently shown promise for rapid AST but challenges of bacterial immobilization can lead to variable results. Herein, a novel cantilever-based method is described for detecting phenotypic antibiotic resistance within ∼45 min, capable of detecting single bacteria. This method does not require complex, variable bacterial immobilization and instead uses a laser and detector system to detect single bacterial cells in media as they pass through the laser focus. This provides a simple readout of bacterial antibiotic resistance by detecting growth (resistant) or death (sensitive), much faster than the current methods. The potential of this technique is demonstrated by determining the resistance in both laboratory and clinical strains of Escherichia coli (E. coli), a key species responsible for clinically burdensome urinary tract infections. This work provides the basis for a simple and fast diagnostic tool to detect antibiotic resistance in bacteria, reducing the health and economic burdens of AMR.In this work, a unique comprehensive and comparative analysis of photoinduced charge generation, recombination kinetics, and energy losses has been carried out to study the effect of different fullerene-based acceptors (FBAs) and nonfullerene acceptors (NFAs) on the performance of organic solar cells (OSCs). For this, different FBAs, specifically ICBA, PC60BM, and PC70BM, and NFAs, namely, ITIC, IT-4F, and IEICO-4F, were employed independently along with a particular donor polymer, PBDB-T, to fabricate bulk heterojunction OSCs and their performances have been compared. This donor molecule is known to give similar power conversion efficiency (PCE) with FBAs and NFAs and hence is ideal for comparative studies. The origin of the higher PCE of NFA-based OSCs vs FBA-based OSCs is analyzed in terms of spectral coverage, charge generation, recombination, and energy loss. It is found that the energy loss (ΔEloss) is ∼0.8 to 1 eV for FBA-based OSCs, while it is 0.5-0.7 eV for NFA-based OSCs. Interestingly, for the PBDB-TIEICO-4F-based system, energy losses due to charge generation (ΔECT) are ∼0 eV and therefore this system has minimum ΔEloss among all of the studied devices. Providing a systematic, comprehensive, and comparative outlook, our study may further be extended to new upcoming NFA systems and beyond the donor system used in this work.Fluorescence methods are important tools to identify RNA-binding small molecules and further employed to study RNA-protein interactions. Most reported fluorescence strategies are based on covalent labeling of ligand or RNA, which can impede the binding between them to some extent, or light-off fluorescent indicator displacement methods, which ask for particular indicators. Herein, a label-free fluorescence strategy based on the light-on aggregation-induced emission (AIE) feature of tetraphenylethene (TPE) derivative to screen RNA-binding small molecules is presented. As a result of electrostatic interaction, the selected peptides can induce self-assembly of the TPE derivative to produce strong fluorescent emission; when the peptides are bound to RNA molecules, the TPE derivative is in the deaggregated form and shows no or minimum fluorescence. Based on the phenomenon, a competitive displacement assay combined with the TPE reporter was employed to characterize selected small molecules for their binding abilities to HIV-I RNAs. This AIE feature enables the fluorescence-off state of the TPE derivative in the presence of RNA-peptide complex to be "lightened up" quickly as the RNA-binding molecule is introduced and the peptide is competitively released. This strategy was carried out to test several small molecule binders, and the results are consistent with previous reports. This report gives an inspiring example of AIE-based fluorescent assay for HIV-I RNA-binding molecule screening, which may further be explored to build a drug screening platform for RNA-protein interference.Drug-induced hepatic damage has drawn great attention on public health problems. Drugs are biotransformed in the liver by enzymatic processes, accompanied by the production of reactive free radicals, which is the main cause of drug-induced hepatotoxicity. However, the limited penetration of optics makes the use of current luminescence imaging more difficult for acquiring free radicals mapping for lesion location, when applied to whole-body imaging in vivo. In this work, we develop an activatable nanoprobe based on Prussian blue (PB) that can combine magnetic resonance imaging (MRI) and photoacoustic imaging (PAI) for deep-tissue ONOO- imaging. We discover that ONOO- can oxidize FeII within PB into FeIII and meanwhile destroy the crystal structure of PB so that the strong absorption of PB at 710 nm that originated from the electron transferring between FeII and FeIII is greatly diminished. BI-3406 inhibitor As a result, the reduced photoacoustic imaging (PA) signal of PB is able to function as an indicator for sensing ONOO-. Importantly, after reaction with ONOO-, the reduced size of PB results in the decrease of rotational correlation time (τR), leading to the activatable MRI signal for sensing ONOO-.
Homepage: https://www.selleckchem.com/products/bi-3406.html
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