Optimizing multi-dimensional high throughput screening using zebrafish

- Choice of chemical delivery method affects the bioactivity profile.
- Digital dispensing results in more accurate chemical delivery.
- Extensive mixing increases chemical dispersion apparent bioactivity.
- Zebrafish model is sensitive to chemical impurities.

The use of zebrafish for high throughput screening (HTS) for chemical bioactivity assessments is becoming routine in the fields of drug discovery and toxicology. Here we report current recommendations from our experiences in zebrafish HTS. We compared the effects of different high throughput chemical delivery methods on nominal water concentration, chemical sorption to multi-well polystyrene plates, transcription responses, and resulting whole animal responses. We demonstrate that digital dispensing consistently yields higher data quality and reproducibility compared to standard plastic tip-based liquid handling. Additionally, we illustrate the challenges in using this sensitive model for chemical assessment when test chemicals have trace impurities. Adaptation of these better practices for zebrafish HTS should increase reproducibility across laboratories.