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Objective The purpose of this experiment is to monitor the effects of different concentrations of LiCl on anterior development in zebrafish embryos. This will be carried out by assessing the degree to which certain amounts of the lithium cholride teratogen affect the morphology of anterior structures such as the eye. Materials
Procedure 1. Obtain 15-20 zebrafish embryos at the sphere/dome-stage of development (four hours post-fertilization) for each of the four titrations of lithium chloride to be tested, inclusive of the control group. Therefore, about 60-80 sphere/dome-stage embryos must be isolated in total. For a description of sphere/dome-stage embryos, refer to the Zebrafish Information Network homepage (www.zfin.org). 2. Place the embryos into four separate petri dishes, each containing 10 mL of Zebrafish embryo medium. 3. Next, procure three dishes and fill them with the following concentrations of LiCl:
A table including the calculations for the three LiCl titrations used in the original experiment is included in the Materials and Methods section of this page. 4. Transfer each population of embryos from the embryo medium to the LiCl-containing dishes using a severed pipette and immerse them in solution for 10 minutes. 5. After 10 minutes, rinse the embryos by placing each group in a separate dish containing 10 milliliters of embryo medium, and then transfer back to the original embryo-medium-containing dishes. Be sure to label the dishes with the corresponding amount of LiCl to which they were induced. 6. Photograph the embryos after 24 hours to observe anterior development, carefully noting deviations in anterior patterning and formation from the control population.
J. N. White 5/13/04 |
