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Novel droplet-based approach for investigating bacterial biofilm formation on microplastic
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Researchers developed a droplet-based microfluidic approach to study bacterial biofilm formation on microplastic surfaces, enabling high-throughput screening of how different polymer types and surface conditions influence plastisphere community development.
One of several key threats associated with the increasing global plastic pollution is plastics ability to aid microorganisms in biofilm formation [1][2]. Biofilms promote development of antimicrobial resistance (AMR), one of the most pressing health concerns in our society [3]. However, despite the urgency, there are still many knowledge gaps. One such aspect still poorly understood is how microplastic effects biofilm formation [4]. This is due to the difficulty of studying such small particles [4]. Using droplet-based technology could provide an excellent solution, as it allows compartmentalization of study material into hundreds of thousands of nano-and picolitre droplets that can be incubated and analyzed via different methods [5]. We show that our user-friendly droplet-based emulsion pipeline can be used to investigate microplastic modulation of AMR and potential biofilm formation. We generated monodisperse water-in-oil droplets with using a poly(dimethylsiloxane) (PDMS) microfluidic chip with flow-focusing geometry. We performed single cell-based (droplet) minimal inhibitory concentration (MIC) assays with GFP-labelled Escherichia coli and antibiotics Cefotaxime and Kanamycin respectively. Each droplet MIC assay contained samples with and without added 10 µm carboxylated polystyrene microspheres. Droplets were incubated for 24h at 37oC, followed by fluorescence imaging of droplet monolayer and analysis via software Ilastik [6] and CellProfiler[7]. Results showed that our droplet-based emulsion pipeline enables (i) identifying microplastic inside nanoliter droplets, (ii) studying potential antibiotic specific susceptibility of bacteria, and (iii) investigate possible early biofilm formation (autoaggragation of bacteria) trends. Our pipeline moreover allowed conducting experiments with thousands of parallel replicates. References [1] doi: 10.1007/s44169-023-00035-z. [2] doi: 10.1016/j.hazadv.2022.100077. [3] doi: 10.1371/journal.pmed.1004264 [4] doi: 10.3389/FMICB.2021.603967/BIBTEX. [5] doi: 10.1021/acs.analchem.9b05047. [6] doi: 10.1038/s41592-019-0582-9. [7] doi: 10.1186/gb-2006-7-10-r100. Also see: https://micro2024.sciencesconf.org/553221/document
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