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Title: Analysis of Factors Limiting Bacterial Growth in PDMS Mother Machine Devices

Abstract

The microfluidic mother machine platform has attracted much interest for its potential in studies of bacterial physiology, cellular organization, and cell mechanics. Despite numerous experiments and development of dedicated analysis software, differences in bacterial growth and morphology in narrow mother machine channels compared to typical liquid media conditions have not been systematically characterized. Here we determine changes in E. coli growth rates and cell dimensions in different sized dead-end microfluidic channels using high resolution optical microscopy. We find that E. coli adapt to the confined channel environment by becoming narrower and longer compared to the same strain grown in liquid culture. Cell dimensions decrease as the channel length increases and width decreases. These changes are accompanied by increases in doubling times in agreement with the universal growth law. In channels 100 μm and longer, cell doublings can completely stop as a result of frictional forces that oppose cell elongation. Before complete cessation of elongation, mechanical stresses lead to substantial deformation of cells and changes in their morphology. Lastly, our work shows that mechanical forces rather than nutrient limitation are the main growth limiting factor for bacterial growth in long and narrow channels.

Authors:
 [1];  [1];  [2]; ORCiD logo [3];  [1]
  1. The Univ. of Tennessee, Knoxville, TN (United States)
  2. Univ. of Tennessee, Knoxville, TN (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1436023
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Microbiology
Additional Journal Information:
Journal Volume: 9; Journal ID: ISSN 1664-302X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; mother machine; nutrient shielding; mechanics of cell growth; peptidoglycan synthesis; cell wall; microfluidics

Citation Formats

Yang, Da, Jennings, Anna D., Borrego, Evalynn, Retterer, Scott T., and Mannik, Jaan. Analysis of Factors Limiting Bacterial Growth in PDMS Mother Machine Devices. United States: N. p., 2018. Web. doi:10.3389/fmicb.2018.00871.
Yang, Da, Jennings, Anna D., Borrego, Evalynn, Retterer, Scott T., & Mannik, Jaan. Analysis of Factors Limiting Bacterial Growth in PDMS Mother Machine Devices. United States. doi:10.3389/fmicb.2018.00871.
Yang, Da, Jennings, Anna D., Borrego, Evalynn, Retterer, Scott T., and Mannik, Jaan. Tue . "Analysis of Factors Limiting Bacterial Growth in PDMS Mother Machine Devices". United States. doi:10.3389/fmicb.2018.00871. https://www.osti.gov/servlets/purl/1436023.
@article{osti_1436023,
title = {Analysis of Factors Limiting Bacterial Growth in PDMS Mother Machine Devices},
author = {Yang, Da and Jennings, Anna D. and Borrego, Evalynn and Retterer, Scott T. and Mannik, Jaan},
abstractNote = {The microfluidic mother machine platform has attracted much interest for its potential in studies of bacterial physiology, cellular organization, and cell mechanics. Despite numerous experiments and development of dedicated analysis software, differences in bacterial growth and morphology in narrow mother machine channels compared to typical liquid media conditions have not been systematically characterized. Here we determine changes in E. coli growth rates and cell dimensions in different sized dead-end microfluidic channels using high resolution optical microscopy. We find that E. coli adapt to the confined channel environment by becoming narrower and longer compared to the same strain grown in liquid culture. Cell dimensions decrease as the channel length increases and width decreases. These changes are accompanied by increases in doubling times in agreement with the universal growth law. In channels 100 μm and longer, cell doublings can completely stop as a result of frictional forces that oppose cell elongation. Before complete cessation of elongation, mechanical stresses lead to substantial deformation of cells and changes in their morphology. Lastly, our work shows that mechanical forces rather than nutrient limitation are the main growth limiting factor for bacterial growth in long and narrow channels.},
doi = {10.3389/fmicb.2018.00871},
journal = {Frontiers in Microbiology},
number = ,
volume = 9,
place = {United States},
year = {2018},
month = {5}
}

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