High-resolution imaging and analysis of the intestinal bacterial load of<i>Caenorhabditis elegans</i>during early adulthood

Rezaeianaran, Farzad; Gijs, Martin A. M.

doi:10.1039/d3ra02934d

Cited by 1 publication

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“…This observation prompts us to employ different methods for the analysis of the bacterial load between the fluorescent E. coli -OP50-fed worms and the fluorescent P. aeruginosa-PAO1-fed ones. In a previous work [ 54 ], we expounded the detection and analysis of fluorescent spots in IMARIS. As the procedure is applicable here, but only to the fluorescent E. coli -OP50-fed worms ( Figure 5 a), we also briefly explain it here.…”

Section: Methodsmentioning

confidence: 99%

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

Rezaeianaran,

Gijs

2023

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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 (60×). 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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Section: Methodsmentioning

confidence: 99%

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

Rezaeianaran,

Gijs

2023

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High-resolution imaging and analysis of the intestinal bacterial load ofCaenorhabditis elegansduring early adulthood

Abstract: 3D representation and quantitative analysis of intestinal bacteria in C. elegans worms.

Cited by 1 publication

References 39 publications

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

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

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