Embryo-Based Chemical Toxicity Screen: Assessing Effects on Developing Zebrafish Embryos
Published: April 30, 2023
Abstract
Source: Hao J. et al., Large Scale Zebrafish-Based In vivo Small Molecule Screen. J. Vis. Exp. (2010)
In this article, zebrafish embryos are used for chemical toxicity screening. Chemical toxicity is correlated with phenotype readouts. The sample protocol shows a methodological approach of performing the chemical toxicity screening.
Protocol
1. Transfer of Small Molecule Library While compound transfer can be automated with robotic transfer methods, we will describe the manual transfer method. Small molecule libraries are typically supplied in a 96-well format, with each compound stored in DMSO as a 10 mM stock. About 60 minutes before the embryos reach the stage when the compounds are to be added, thaw a desired number of 96-well plates containing aliquots of small molecules (source plate). Take note …
Materials
| Small molecule library of structurally diverse compounds arrayed in a 96-wll format at 10 mM stock in DMSO. Each master plate is aliquoted into 96-well polypropylene storage plates (Corning), and stored at -80°C until use. |
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| Drierite | W.A. HAMMOND DRIERITE CO, Xenia, OH | ||
| 12-channel pipettes, 2-20 μL | Eppendorf | ||
| Aluminum sealing tape for 96-well plates | Nunc, Rochester, NY | ||
| DMSO | Sigma, St. Louis, MO | ||
| Basic incubator, 28.5°C | Fisher Scientific | ||
| Polystyrene 96-well round-bottom assay plates | Corning COSTAR; Lowell, MA | ||
| Stereomicroscope with transmitted light base | Leica Microsystems, Bannockburn, IL | ||
| E3 embryo medium: 5 mM NaCl, 0.17 mM KCl, 0.33 mM CaCl2 , 0.33 mM MgSO4 , containing 0.003% PTU (phenylthiocarbamide, Sigma; St. Louis, MO). | PTU can be prepared as a 10x solution by dissolving 0.3-g PTU in 1 L of E3 embryo media. Solutions containing PTU should be protected from light by covering with aluminum foil. |
Bu Makaleden Alıntı Yapın
Embryo-Based Chemical Toxicity Screen: Assessing Effects on Developing Zebrafish Embryos. J. Vis. Exp. (Pending Publication), e20179, doi: (2023).