High-resolution imaging and analysis of the intestinal bacterial load of during early adulthood.

RSC Adv

Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne CH-1015 Lausanne Switzerland

Published: June 2023

We study the presence within the worm () of a fluorescent strain of the worm's bacterial food ( () OP50) during early adulthood. Use of a microfluidic chip based on a thin glass coverslip substrate allows investigation of the intestinal bacterial load using a Spinning Disk Confocal Microscope (SDCM) equipped with a high-resolution objective (60×). High-resolution z-stack fluorescence images of the gut bacteria in adult worms, which were loaded in the microfluidic chip and subsequently fixed, were analyzed using IMARIS software and 3D reconstructions of the intestinal bacterial load in the worms were obtained. We present an automated bivariate histogram analysis of the volumes and intensities of the bacterial spots for each worm and find that, as the worms age, the bacterial load in their hindguts increases. We show the advantage of single-worm resolution automated analysis for bacterial load studies and anticipate that the methods described in our work can be easily implemented in existing microfluidic solutions to enable thorough studies of bacterial proliferation.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10248764PMC
http://dx.doi.org/10.1039/d3ra02934dDOI Listing

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