Counting Caenorhabditis elegans: Protocol Optimization and Applications for Population Growth and Toxicity Studies in Liquid Medium.

摘要: The nematode Caenorhabditis elegans is used extensively in molecular, toxicological and genetics research. However, standardized methods for counting nematodes liquid culture do not exist despite the wide use of need accurate measurements. Herein, we provide a simple affordable protocol developed to maximize count accuracy minimize variability culture. Sources process were identified tested 14 separate experiments. Three variables resulted significant effects on count: shaking culture, priming pipette tips, sampling location within microcentrifuge tube. Between-operator did have statistically effect counts, even among differently-skilled operators. was assess population growth rates two different but common media: axenic modified Habitation Reproduction medium (mCeHR) S-basal complete. In mCeHR, populations doubled daily 10 d. complete initially every 12 h, slowed within 7 We also detected difference between embryo-to-hatchling incubation period 5 d mCeHR compared 4 method reduces allows rigorous reliable experimentation.