Systematic Evaluation of the Effects of Toxicant Exposure on Survival in Zebrafish Embryos and Larvae

Anjana Ramdas Nair, Patrice Delaney, Abigail Ama Koomson, Shashi Ranjan, Kirsten C. Sadler

Research output: Contribution to journalArticlepeer-review


The use of whole animal models in toxicological studies is essential for understanding the physiological responses caused by chemical exposures. However, such studies can face reproducibility challenges due to unaccounted experimental parameters that can have a marked influence on toxicological outcomes. Zebrafish embryos and larvae are a popular vertebrate animal model for studying cellular, tissue, and organ responses to toxicant exposures. Despite the popularity of this system, standardized protocols that control for the influence of various experimental parameters and culture conditions on the toxicological response in these animals have not been widely adopted, making it difficult to compare findings from different laboratories. Here, we describe a detailed approach for designing and optimizing protocols to assess the impact of chemical exposures on the development and survival of zebrafish embryos and larvae. We first describe our standard procedure to determine two key toxicological thresholds, the maximum tolerable concentration (MTC) and the lethal concentration (LC50, defined as that in which 50% of larvae die), in response to an exposure that persists from early development through larval maturation. We then describe two protocols to systematically test how key experimental parameters, including genetic background, culture media, animal density, volume, plate material, and developmental stage in which the embryos are exposed, alter the MTC and LC50. Finally, we provide a step-by-step guide to assess the interaction between two chemicals using this model. These protocols will guide the standardization of toxicological studies using zebrafish and maximize reproducibility.

Original languageEnglish (US)
Article numbere231
JournalCurrent Protocols
Issue number9
StatePublished - Sep 2021


  • pharmacology
  • toxicant exposure
  • toxicology
  • zebrafish embryo
  • zebrafish larvae
  • Water Pollutants, Chemical
  • Reproducibility of Results
  • Animals
  • Larva
  • Zebrafish
  • Lethal Dose 50

ASJC Scopus subject areas

  • Medical Laboratory Technology
  • General Medicine
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • Health Informatics
  • General Neuroscience
  • Pharmacology, Toxicology and Pharmaceutics(all)


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