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Cell Biology JoVE (Open Access) Citable · DOI

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans

DOI: 10.3791/56892-v
What you'll learn
  • ✓ Prepare synchronized C. elegans cultures in 96-well plates for high-throughput screening
  • ✓ Apply chemical treatments to nematode populations and assess phenotypic responses
  • ✓ Execute and validate compound screening assays for longevity effects
Protocol

Here we describe a simple protocol for rapidly producing hundreds of nematode growth media agar, 96-well culture plates with consistent numbers of Caenorhhabditis elegans per well. These cultures are useful for the phenotypic screening of whole organisms. We focus here on using these cultures to screen chemicals for pro-longevity effects.

Difficulty
beginner
Total time
~3–5 days (including culture synchronization and assay incubation)
Model organism
Caenorhabditis elegans
Biosafety
BSL-1

Steps

1
Prepare NGM plates and mass culture C. elegans

Pour nematode growth medium (NGM) agar plates and establish a bulk C. elegans culture to generate sufficient synchronized worms for downstream screening.

▶ 00:30
2
Hatch synchronized population of arrested larvae

Use buffer-based hatching to obtain a synchronized culture of L1-arrested larvae, ensuring uniform developmental stage across all experimental wells.

▶ 03:24
3
Setup and treat 96-well culture plates with chemicals

Distribute synchronized L1 larvae into 96-well plates at consistent density and apply chemical treatments to test compounds for phenotypic effects.

▶ 05:01
4
Analyze screening results and validate hit compounds

Assess phenotypic outcomes from initial compound screening, identify candidate compounds, and perform re-testing to confirm reproducibility and pro-longevity effects.

▶ 06:42
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