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

Abstract

The nematode Caenorhabditis elegans has been extensively used as a model multicellular organism to study the influence of osmotic stress conditions and the toxicity of chemical compounds on developmental and motilityassociated phenotypes. However, the several-day culture of nematodes needed for such studies has caused researchers to explore alternatives. In particular, C. elegans embryos, due to their shorter developmental time and immobile nature, could be exploited for this purpose, although usually their harvesting and handling is tedious. Here, we present a multiplexed, high-throughput and automated embryo phenotyping microfluidic approach to observe C. elegans embryogenesis after the application of different chemical compounds. After performing experiments with up to 800 embryos per chip and up to 12 h of time-lapsed imaging per embryo, the individual phenotypic developmental data were collected and analyzed through machine learning and image processing approaches. Our proof-of-concept platform indicates developmental lag and the induction of mitochondrial stress in embryos exposed to high doses (200 mM) of glucose and NaCl, while small doses of sucrose and glucose were shown to accelerate development. Overall, our new technique has potential for use in large-scale developmental biology studies and opens new avenues for very rapid high-throughput and high-content screening using C. elegans embryos.

Non-Rodent Genetic Animal Models for Studying Tauopathy: Review of Drosophila, Zebrafish, and C. elegans Models

“…Generally, behavioral phenotype assays to test Tau toxicity in C. elegans are mobilitybased [233]. In the thrashing assay, a well-established motility-based assay, the frequency and lateral swimming movements of worms placed in liquid media are measured [234].…”

C. elegans: A biosensor for host–microbe interactions

“…through a new set of hardware and software tools that allow the manipulation, recording and imaging of worms grown in hundreds of conditions in parallel. These include the use of flow cytometry-like stations as COPAS 98 to measure worm physical variables such as axial length and optical density; the 'WorMotel' from Fang-Yen 99 and the Lifespan Machine from Fontana 100 that measure lifespan; diverse tracking platforms from Brown 96,101 , Nollen 102 , Rex 103 , Driscoll 104 or Goodman 105 labs that can assess motor-related phenotypical variables including behavior; or fully automated workstations from Pincus 106 or Lu 97 that monitor long-term behavior and healthspan, among many other examples [107][108][109][110] .…”

“…Exploring the transparent nature of C. elegans, fluorescence imaging can also be performed in a high-throughput manner, further extending the screening capabilities of the worm as a biosensor. Fluorescence information can be obtained at different scales, from studying protein dynamics in whole animals 108,112 to processes in particular specific cell lineages 113 . Hence, by using worm molecular or physiological phenotypes as readouts of bacterial activity, it is possible to capture in fine detail several molecular mechanisms at the host level involved in these complex host-microbe interactions.…”

Caenorhabditis elegans for rare disease modeling and drug discovery: strategies and strengths

“…Although both Drosophila and zebrafish provide apt systems for high-content screens, they do not match the ability of C. elegans for high-throughput analysis (Patten et al, 2016). This fact is further illustrated by the advent of multiple protocols to perform such assays, including the more recent use of microfluidic platforms to improve high-throughput screening (Atakan et al, 2020;Kinser and Pincus, 2017;Lucanic et al, 2018;O'Rourke et al, 2009). The utility of C. elegans for rare disease modeling is underscored by their inclusion in model organism screening centers (MOSCs), a component of the newly expanded Undiagnosed Disease Network (UDN; https://undiagnosed.hms.harvard.edu/).…”