Counter-on-chip for bacterial cell quantification, growth, and live-dead estimations.

Autor: Rahman KMT; Department of Biology and Microbiology, South Dakota State University, Brookings, SD, 57006, USA., Butzin NC; Department of Biology and Microbiology, South Dakota State University, Brookings, SD, 57006, USA. nicholas.butzin@gmail.com.; Department of Chemistry, Biochemistry and Physics, South Dakota State University, Brookings, SD, 57006, USA. nicholas.butzin@gmail.com.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Jan 08; Vol. 14 (1), pp. 782. Date of Electronic Publication: 2024 Jan 08.
DOI: 10.1038/s41598-023-51014-2
Abstrakt: Quantifying bacterial cell numbers is crucial for experimental assessment and reproducibility, but the current technologies have limitations. The commonly used colony forming units (CFU) method causes a time delay in determining the actual numbers. Manual microscope counts are often error-prone for submicron bacteria. Automated systems are costly, require specialized knowledge, and are erroneous when counting smaller bacteria. In this study, we took a different approach by constructing three sequential generations (G1, G2, and G3) of counter-on-chip that accurately and timely count small particles and/or bacterial cells. We employed 2-photon polymerization (2PP) fabrication technology; and optimized the printing and molding process to produce high-quality, reproducible, accurate, and efficient counters. Our straightforward and refined methodology has shown itself to be highly effective in fabricating structures, allowing for the rapid construction of polydimethylsiloxane (PDMS)-based microfluidic devices. The G1 comprises three counting chambers with a depth of 20 µm, which showed accurate counting of 1 µm and 5 µm microbeads. G2 and G3 have eight counting chambers with depths of 20 µm and 5 µm, respectively, and can quickly and precisely count Escherichia coli cells. These systems are reusable, accurate, and easy to use (compared to CFU/ml). The G3 device can give (1) accurate bacterial counts, (2) serve as a growth chamber for bacteria, and (3) allow for live/dead bacterial cell estimates using staining kits or growth assay activities (live imaging, cell tracking, and counting). We made these devices out of necessity; we know no device on the market that encompasses all these features.
(© 2024. The Author(s).)
Databáze: MEDLINE
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