Journal
LAB ON A CHIP
Volume 20, Issue 1, Pages 54-63Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c9lc00684b
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Funding
- European Union [711859]
- Polish Ministry of Science and Higher Education
- National Science Centre grant [DEC-2014/12/W/NZ6/00454]
- National Science Centre, Poland, within the project Sonata [2016/21/D/ST3/00988]
- Foundation for Polish Science within the Team-Tech/2016-2/10 program
- Ministry of Science and Higher Education [0722/E 64/STYP/10/295]
- NanoFun [POIG.02.02.00-00-025/09]
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The alarming dynamics of antibiotic-resistant infections calls for the development of rapid and point-of-care (POC) antibiotic susceptibility testing (AST) methods. Here, we demonstrated the first completely stand-alone microfluidic system that allowed the execution of digital enumeration of bacteria and digital antibiograms without any specialized microfluidic instrumentation. A four-chamber gravity-driven step emulsification device generated similar to 2000 monodisperse 2 nanoliter droplets with a coefficient of variation of 8.9% of volumes for 95% of droplets within less than 10 minutes. The manual workload required for droplet generation was limited to the sample preparation, the deposition into the sample inlet of the chip and subsequent orientation of the chip vertically without an additional pumping system. The use of shallow chambers imposing a 2D droplet arrangement provided superior stability of the droplets against coalescence and minimized the leakage of the reporter viability dye between adjacent droplets during long-term culture. By using resazurin as an indicator of the growth of bacteria, we were also able to reduce the assay time to similar to 5 hours compared to 20 hours using the standard culture-based test.
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