Rapid, high-throughput tracking of bacterial motility in 3D via phase-contrast holographic video microscopy

Biophysical Journal
Fook Chiong CheongChwee Teck Lim

Abstract

Tracking fast-swimming bacteria in three dimensions can be extremely challenging with current optical techniques and a microscopic approach that can rapidly acquire volumetric information is required. Here, we introduce phase-contrast holographic video microscopy as a solution for the simultaneous tracking of multiple fast moving cells in three dimensions. This technique uses interference patterns formed between the scattered and the incident field to infer the three-dimensional (3D) position and size of bacteria. Using this optical approach, motility dynamics of multiple bacteria in three dimensions, such as speed and turn angles, can be obtained within minutes. We demonstrated the feasibility of this method by effectively tracking multiple bacteria species, including Escherichia coli, Agrobacterium tumefaciens, and Pseudomonas aeruginosa. In addition, we combined our fast 3D imaging technique with a microfluidic device to present an example of a drug/chemical assay to study effects on bacterial motility.

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Citations

Nov 3, 2015·Seminars in Cell & Developmental Biology·Timothy Krüger, Markus Engstler
Aug 11, 2016·Biophysical Journal·Linda TurnerHoward C Berg
Mar 17, 2017·Langmuir : the ACS Journal of Surfaces and Colloids·Meng QiGuangzhao Zhang
Nov 3, 2015·Nature Communications·K M TauteT S Shimizu
Mar 6, 2019·The Review of Scientific Instruments·James L FlewellenRichard M Berry
May 6, 2016·Micromachines·Yaxiaer Yalikun, Yo Tanaka
Nov 13, 2017·Optics Express·Nicola E FarthingLaurence G Wilson
Aug 25, 2017·ACS Nano·Jaime de AndaGerard C L Wong

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Methods Mentioned

BETA
confocal microscopy
imaging technique

Software Mentioned

IDL

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