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The conventional antifungal screening techniques, such as water agar and 96-well plates, are based on laborious protocols and bulk analysis, restricting the analysis at the single spore level and are time consuming. In this study, we present a droplet-based microfluidic platform that enables antifungal analysis of single spores of filamentous fungus <jats:italic>Alternaria alternata<\/jats:italic>. A droplet-based viability assay was developed, allowing the germination and hyphal growth of single <jats:italic>A. alternata<\/jats:italic> spores within droplets. The viability was demonstrated over a period of 24\u00a0h and the antifungal screening was achieved using Kunshi\/Tezuma as antifungal agent. The efficacy results of the droplet-based antifungal analysis were compared and validated with the results obtained from conventional protocols. The percentage inhibitions assessed by the droplet-based platform were equivalent with those obtained by the other two methods, and the Pearson correlation analysis showed high correlation between the three assays. Taken together, this droplet-based microfluidic platform provides a wide range of potential applications for the analysis of fungicide resistance development as well as combinatorial screening of other antimicrobial agents and even antagonistic fungi.<\/jats:p>","DOI":"10.1038\/s41598-021-02350-8","type":"journal-article","created":{"date-parts":[[2021,11,26]],"date-time":"2021-11-26T11:02:47Z","timestamp":1637924567000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Droplet-based microfluidics platform for antifungal analysis against filamentous fungi"],"prefix":"10.1038","volume":"11","author":[{"given":"Sehrish","family":"Iftikhar","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aur\u00e9lie","family":"Vigne","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia Elisa","family":"Sepulveda-Diaz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,11,26]]},"reference":[{"key":"2350_CR1","first-page":"32","volume":"56","author":"P Nolte","year":"2015","unstructured":"Nolte, P. 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