Protocol for the Preparation of Zebrafish Embryos for LLO Analysis 1) 150-200 4-7 day old embryos are collected in an 1.8 mL eppendorf tube and all excess water is removed (if using 4 day old or younger embryos, yolks are removed prior to preparation) 2) 1 mL of methanol at room temperature is added to the tube and followed by brief sonication (low setting, 3 x 5 sec pulses). 3) This mixture is then transferred into a 15-mL conical tube and an additional 10-12 mL of methanol is added. 4) Following thorough vortexing, the tube is sealed and sent at room temperature.