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Procedure
Embryo Harvesting
1) Induce adult zebrafish to mate at the beginning of their light cycle by
covering the bottom of the aquarium with a layer of marbles the night before.
This will both excite the fish and protect the embryos from predation. Leave
overnight.
2) Harvest the embryos between one and three hours after the fish have awakened
(at the appearance of light) by using a suction vacuum to clean the bottom,
removing embryos from under the marbles into a filter.
3) Wash embryos from filter into a dish of aquarium water. Sort the embryos
from extraneous debris by using a wide-mouth pipet to remove them into a Petri
dish of ZE solution.
4) Stage the embryos and allow to develop to desired stage.
Solution Preparation
1) Dilute arsenic to desired concentrations (preliminary assay should
be performed to determine effective range) using extreme caution
at high concentrations, as arsenic is highly toxic. The earliest
dilutions may be made with distilled water; later solutions should
be diluted in ZE solution.
2) Put approximately 10 mL of solutions into labeled Petri dishes.
Make a control solution with 10 mL ZE solution.
2) Carefully clean or dispose of pipets and other exposed equipment.
Embryo Treatment
1) Divide embryos between the prepared solutions. Cover the dishes to prevent
excessive evaporation.
2) Place in incubator and allow to develop.
Embryo Observation
1) Examine embryos under the dissecting microscope approximately 24 hours after
fertilization.
2) Stage embryos and note any deformities observed.
3) Photograph selected embryos to document results.
4) Observe embryos at periodic intervals thereafter and document results.
  
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