Dataset Information

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

ABSTRACT: 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.

SUBMITTER: Rahman KMT

PROVIDER: S-EPMC10774380 | biostudies-literature | 2024 Jan

REPOSITORIES: biostudies-literature

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

Rahman K M Taufiqur KMT Butzin Nicholas C NC

Scientific reports 20240108 1

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 seq ...[more]

PMID: 38191788

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Project description:Nanotopographic surfaces are proven to be successful in killing bacterial cells upon contact. This non-chemical bactericidal property has paved an alternative way of fighting bacterial colonization and associated problems, especially the issue of bacteria evolving resistance against antibiotic and antiseptic agents. Recent advancements in nanotopographic bactericidal surfaces have made them suitable for many applications in medical and industrial sectors. The bactericidal effect of nanotopographic surfaces is classically studied under static conditions, but the actual potential applications do have fluid flow in them. In this study, we have studied how fluid flow can affect the adherence of bacterial cells on nanotopographic surfaces. Gram-positive and Gram-negative bacterial species were tested under varying fluid flow rates for their retention and viability after flow exposure. The total number of adherent cells for both species was reduced in the presence of flow, but there was no flowrate dependency. There was a significant reduction in the number of live cells remaining on nanotopographic surfaces with an increasing flowrate for both species. Conversely, we observed a flowrate-independent increase in the number of adherent dead cells. Our results indicated that the presence of flow differentially affected the adherent live and dead bacterial cells on nanotopographic surfaces. This could be because dead bacterial cells were physically pierced by the nano-features, whereas live cells adhered via physiochemical interactions with the surface. Therefore, fluid shear was insufficient to overcome adhesion forces between the surface and dead cells. Furthermore, hydrodynamic forces due to the flow can cause more planktonic and detached live cells to collide with nano-features on the surface, causing more cells to lyse. These results show that nanotopographic surfaces do not have self-cleaning ability as opposed to natural bactericidal nanotopographic surfaces, and nanotopographic surfaces tend to perform better under flow conditions. These findings are highly useful for developing and optimizing nanotopographic surfaces for medical and industrial applications.

Dataset Information

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

Publications

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

Similar Datasets

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