Automated high-content phenotyping from the first larval stage till the onset of adulthood of the nematodeCaenorhabditis elegans

Abstract

The nematode Caenorhabditis elegans is increasingly used as a model for human biology. However, in vivo culturing platforms for C. elegans allowing high-content phenotyping during their life cycle in an automated fashion are lacking so far. Here, a multiplexed microfluidic platform for the rapid high-content phenotyping of populations of C. elegans down to single animal resolution is presented. Nematodes are (i) reversibly and regularly confined during their life inside tapered channels for imaging fluorescent signal expression and to measure their growth parameters and, (ii) allowed to freely move in microfluidic chambers, during which swimming behavior was video-recorded. The obtained data sets are analyzed in an automated way and 19 phenotypic parameters are extracted. Our platform is employed for studying the effect of bacteria dilution, a form of dietary restriction (DR) in nematodes, on a worm model of Huntington disease and demonstrates the influence of DR on disease regression.

Automated Platform for Long-Term Culture and High-Content Phenotyping of Single C. elegans Worms

“…In parallel to the head amplitude study, the tail amplitude behavior up to 10 µM was not significantly different from the control, but at 100 µM and 1 mM tetramisole concentrations, the movement was reduced. Overall, the longitudinal phenotyping of the motility parameters did not clearly highlight an anthelmintic dose influence on the nematodes while proving that the nominal values were within an expected interval 12 . As illustrated here, our platform however brings significant advantages in terms of tracking the whole dynamics of single worms under a chemical exposure and thus obtaining phenotypic data longitudinally or with a dose-response approach.…”

“…We observed the growth dynamics of N2 wild-type nematodes in their chambers, when exposed to 1 nM to 1 mM tetramisole concentrations. Our automated video processing code, which was modified from a previous work 12,29 , allowed us to accurately quantify longitudinal phenotypes (Supplementary Fig. S5).…”

“…In addition to the larval development parameters, we selected therefore four phenotypic motility parameters, namely, central velocity, head amplitude, beating frequency and tail amplitude 12 , to study the tetramisole dose-effect on C . elegans as a function of development time.…”

“…However, the majority of the proposed C . elegans microfluidic platforms were designed for studying of nematodes at the whole population level, leading to averaged results and omitting the study of any relevant individual response and eventual intra-population heterogeneity 9–12 . High-resolution and high-throughput imaging as enabled by microfluidics was also used for detecting subtle aging phenotypes 13 or for assaying of poly-glutamine aggregates as indicators of disease progression 14 .…”

Automated phenotyping of Caenorhabditis elegans embryos with a high-throughput-screening microfluidic platform

“…The nematode C. elegans is a well-established model organism in terms of rapid data extraction through automated phenotyping studies 26 . Previously, various tracking methods were applied to nematodes to provide high-content phenotypic data [27][28][29] . However, very few automated phenotyping methods have been proposed for C. elegans embryos.…”