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  4. Difference in Intestine Content of Caenorhabditis elegans When Fed on Non-Pathogenic or Pathogenic Bacteria
 
research article

Difference in Intestine Content of Caenorhabditis elegans When Fed on Non-Pathogenic or Pathogenic Bacteria

Rezaeianaran, Farzad  
•
Gijs, Martin A. M.  
July 1, 2023
Micromachines

We investigated the bacterial food digestion and accumulation in wild-type adult Caenorhabditis elegans (C. elegans) worms that have fed on either non-pathogenic RFP-expressing Escherichia coli (E. coli) OP50 or pathogenic-RFP-expressing Pseudomonas aeruginosa (P. aeruginosa) PAO1 during the first 4 days of adulthood. Once the worms had completed their planned feeding cycles, they were loaded on microfluidic chips, where they were fixed to allow high-resolution z-stack fluorescence imaging of their intestines utilizing a Spinning Disk Confocal Microscope (SDCM) equipped with a high-resolution oil-immersion objective (60x). IMARIS software was used to visualize and analyze the obtained images, resulting in the production of three-dimensional constructs of the intestinal bacterial load. We discovered two distinct patterns for the bacteria-derived fluorescence signal in the intestine: (i) individual fluorescent spots, originating from intact bacteria, were present in the fluorescent E. coli-OP50-fed worms, and (ii) individual fluorescent spots (originating from intact bacteria) were dispersed in large regions of diffuse fluorescence (RDF), originating from disrupted bacteria, in fluorescent P. aeruginosa-PAO1-fed worms. We performed a semi-automated single-worm-resolution quantitative analysis of the intestinal bacterial load, which showed that the intestinal bacterial load generally increases with age of the worms, but more rapidly for the fluorescent P. aeruginosa-PAO1-fed worms.

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Type
research article
DOI
10.3390/mi14071386
Web of Science ID

WOS:001039863800001

Author(s)
Rezaeianaran, Farzad  
Gijs, Martin A. M.  
Date Issued

2023-07-01

Publisher

MDPI

Published in
Micromachines
Volume

14

Issue

7

Article Number

1386

Subjects

Chemistry, Analytical

•

Nanoscience & Nanotechnology

•

Instruments & Instrumentation

•

Physics, Applied

•

Chemistry

•

Science & Technology - Other Topics

•

Physics

•

c

•

elegans

•

high-resolution imaging and analysis

•

bacterial load

•

microfluidics

•

pseudomonas-aeruginosa virulence

•

c. elegans

•

microbiota

•

model

•

systems

•

worms

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LMIS2  
Available on Infoscience
August 28, 2023
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/200083
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