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

Published

January 17, 2018

Author(s)

Steven Lund, Sanem Hosbas Coskun, Monique Johnson, Christopher Sims, Elijah Petersen, John T. Elliott, Bryant C. Nelson, Leona D. Scanlan, Shannon K. Hanna, Karina Brignoni, Patricia Lapasett

Abstract

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

Citation

Nature - Scientific Reports

Pub Type

Journals

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Keywords

whole organism metrology, Caenorhabditis elegans, counting, population growth rates, standardization

Citation

Lund, S. , Hosbas Coskun, S. , Johnson, M. , Sims, C. , Petersen, E. , Elliott, J. , Nelson, B. , Scanlan, L. , Hanna, S. , Brignoni, K. and Lapasett, P. (2018), Counting Caenorhabditis elegans: Protocol Optimization and Applications for Population Growth and Toxicity Studies in Liquid Medium, Nature - Scientific Reports, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=923527 (Accessed November 21, 2024)