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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Bibliographic Details
Published in:	Frontiers in microbiology Vol. 13; p. 836808
Main Authors:	Dubay, Megan M, Johnston, Nikki, Wronkiewicz, Mark, Lee, Jake, Lindensmith, Christian A, Nadeau, Jay L
Format:	Journal Article
Language:	English
Published:	Switzerland Frontiers Media S.A 21-04-2022
Subjects:	Bacillus subtilis bacterial motility heat shock holographic microscopy Microbiology temperature effects thermophile bacterial motility holographic microscopy Bacillus subtilis temperature effects heat shock tracking thermophile
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Summary:	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 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 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.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Edited by: Iain G. Duggin, University of Technology Sydney, Australia This article was submitted to Extreme Microbiology, a section of the journal Frontiers in Microbiology Reviewed by: Alex Bisson, Brandeis University, United States; Georgia Squyres, California Institute of Technology, United States
ISSN:	1664-302X 1664-302X
DOI:	10.3389/fmicb.2022.836808