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Biofilm development of an opportunistic model bacterium analysed at high spatiotemporal resolution in the framework of a precise flow cell.

ABSTRACT: Life of bacteria is governed by the physical dimensions of life in microscales, which is dominated by fast diffusion and flow at low Reynolds numbers. Microbial biofilms are structurally and functionally heterogeneous and their development is suggested to be interactively related to their microenvironments. In this study, we were guided by the challenging requirements of precise tools and engineered procedures to achieve reproducible experiments at high spatial and temporal resolutions. Here, we developed a robust precise engineering approach allowing for the quantification of real-time, high-content imaging of biofilm behaviour under well-controlled flow conditions. Through the merging of engineering and microbial ecology, we present a rigorous methodology to quantify biofilm development at resolutions of single micrometre and single minute, using a newly developed flow cell. We designed and fabricated a high-precision flow cell to create defined and reproducible flow conditions. We applied high-content confocal laser scanning microscopy and developed image quantification using a model biofilm of a defined opportunistic strain, Pseudomonas putida OUS82. We observed complex patterns in the early events of biofilm formation, which were followed by total dispersal. These patterns were closely related to the flow conditions. These biofilm behavioural phenomena were found to be highly reproducible, despite the heterogeneous nature of biofilm.

SUBMITTER: Lim CP

PROVIDER: S-EPMC5515269 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature

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Biofilm development of an opportunistic model bacterium analysed at high spatiotemporal resolution in the framework of a precise flow cell.

Lim Chun Ping CP Mai Phuong Nguyen Quoc PNQ Roizman Sade Dan D Lam Yee Cheong YC Cohen Yehuda Y

NPJ biofilms and microbiomes 20161019

Life of bacteria is governed by the physical dimensions of life in microscales, which is dominated by fast diffusion and flow at low Reynolds numbers. Microbial biofilms are structurally and functionally heterogeneous and their development is suggested to be interactively related to their microenvironments. In this study, we were guided by the challenging requirements of precise tools and engineered procedures to achieve reproducible experiments at high spatial and temporal resolutions. Here, we ...[more]

PMID: 28721252

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Project description:To validate a prospectively triggered spiral phase-contrast magnetic resonance (MR) sequence accelerated with sensitivity encoding (SENSE) in a population of children and adults with congenital heart disease.The local research ethics committee approved this study, and written consent was obtained from all patients or guardians. Stroke volumes were quantified in 40 patients (mean age ± standard deviation: 21.4 years ± 13.8, age range: 3.0-61.3 years; 22 male patients aged 3.0-38.0 years [mean age, 17.2 years ± 10.5], 18 female patients aged 4.7-61.3 years [mean age, 26.6 years ± 15.9]) with congenital heart disease in the aorta (n = 40), main pulmonary artery (n = 38), right pulmonary artery (n = 22), and left pulmonary artery (n = 24). Stroke volumes were obtained with (a) breath-hold spiral phase-contrast MR imaging with SENSE, (b) conventional breath-hold cartesian phase-contrast MR imaging, and (c) reference free-breathing phase-contrast MR imaging. Stroke volumes were compared by using repeated-measures analysis of variance, Bland-Altman analysis, and correlation coefficients.Imaging time with the breath-hold spiral phase-contrast MR sequence was significantly lower than that with the conventional breath-hold phase-contrast MR sequence (~5 seconds vs ~16 seconds, respectively; P < .0001). There was excellent agreement in stroke volumes in all vessels between the reference free-breathing sequence (mean volume, 60.3 mL ± 27.3) and the two breath-hold sequences-spiral SENSE phase-contrast MR imaging (mean volume, 59.5 mL ± 27.1; P < .001) and conventional cartesian phase-contrast MR imaging (mean volume, 59.8 mL ± 27.6; P = .268). The limits of agreement were smaller with the spiral breath-hold sequence than with the conventional breath-hold sequence (-4.4 mL, 2.9 mL vs -10.3 mL, 9.3 mL, respectively); correlation was similar (r = 0.998 vs r = 0.984, respectively).Flow volumes can be accurately and reliably quantified by using a spiral SENSE phase-contrast MR sequence, with high spatiotemporal resolution obtained in a short breath hold.

Dataset Information

Biofilm development of an opportunistic model bacterium analysed at high spatiotemporal resolution in the framework of a precise flow cell.

Publications

Biofilm development of an opportunistic model bacterium analysed at high spatiotemporal resolution in the framework of a precise flow cell.

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