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Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking

We describe a system for high-temperature investigations of bacterial motility using a digital holographic microscope completely submerged in heated water. Temperatures above 90°C could be achieved, with a constant 5°C offset between the sample temperature and the surrounding water bath. Using this...

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Autores principales: Dubay, Megan M., Johnston, Nikki, Wronkiewicz, Mark, Lee, Jake, Lindensmith, Christian A., Nadeau, Jay L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9069135/
https://www.ncbi.nlm.nih.gov/pubmed/35531296
http://dx.doi.org/10.3389/fmicb.2022.836808
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author Dubay, Megan M.
Johnston, Nikki
Wronkiewicz, Mark
Lee, Jake
Lindensmith, Christian A.
Nadeau, Jay L.
author_facet Dubay, Megan M.
Johnston, Nikki
Wronkiewicz, Mark
Lee, Jake
Lindensmith, Christian A.
Nadeau, Jay L.
author_sort Dubay, Megan M.
collection PubMed
description We describe a system for high-temperature investigations of bacterial motility using a digital holographic microscope completely submerged in heated water. Temperatures above 90°C could be achieved, with a constant 5°C offset between the sample temperature and the surrounding water bath. Using this system, we observed active motility in Bacillus subtilis up to 66°C. As temperatures rose, most cells became immobilized on the surface, but a fraction of cells remained highly motile at distances of >100 μm above the surface. Suspended non-motile cells showed Brownian motion that scaled consistently with temperature and viscosity. A novel open-source automated tracking package was used to obtain 2D tracks of motile cells and quantify motility parameters, showing that swimming speed increased with temperature until ∼40°C, then plateaued. These findings are consistent with the observed heterogeneity of B. subtilis populations, and represent the highest reported temperature for swimming in this species. This technique is a simple, low-cost method for quantifying motility at high temperatures and could be useful for investigation of many different cell types, including thermophilic archaea.
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spelling pubmed-90691352022-05-05 Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking Dubay, Megan M. Johnston, Nikki Wronkiewicz, Mark Lee, Jake Lindensmith, Christian A. Nadeau, Jay L. Front Microbiol Microbiology We describe a system for high-temperature investigations of bacterial motility using a digital holographic microscope completely submerged in heated water. Temperatures above 90°C could be achieved, with a constant 5°C offset between the sample temperature and the surrounding water bath. Using this system, we observed active motility in Bacillus subtilis up to 66°C. As temperatures rose, most cells became immobilized on the surface, but a fraction of cells remained highly motile at distances of >100 μm above the surface. Suspended non-motile cells showed Brownian motion that scaled consistently with temperature and viscosity. A novel open-source automated tracking package was used to obtain 2D tracks of motile cells and quantify motility parameters, showing that swimming speed increased with temperature until ∼40°C, then plateaued. These findings are consistent with the observed heterogeneity of B. subtilis populations, and represent the highest reported temperature for swimming in this species. This technique is a simple, low-cost method for quantifying motility at high temperatures and could be useful for investigation of many different cell types, including thermophilic archaea. Frontiers Media S.A. 2022-04-21 /pmc/articles/PMC9069135/ /pubmed/35531296 http://dx.doi.org/10.3389/fmicb.2022.836808 Text en Copyright © 2022 Dubay, Johnston, Wronkiewicz, Lee, Lindensmith and Nadeau. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Dubay, Megan M.
Johnston, Nikki
Wronkiewicz, Mark
Lee, Jake
Lindensmith, Christian A.
Nadeau, Jay L.
Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title_full Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title_fullStr Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title_full_unstemmed Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title_short Quantification of Motility in Bacillus subtilis at Temperatures Up to 84°C Using a Submersible Volumetric Microscope and Automated Tracking
title_sort quantification of motility in bacillus subtilis at temperatures up to 84°c using a submersible volumetric microscope and automated tracking
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9069135/
https://www.ncbi.nlm.nih.gov/pubmed/35531296
http://dx.doi.org/10.3389/fmicb.2022.836808
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