Lethal Phase - Immunohistochemistry
Wash embryos and place into wash basket.
Dechorionate by pouring bleach directly onto embryos.
Let sit for three minutes.
Rinse embryos in basket with ddH2O until bleach is completely washed away.
Place embryos into glass vial containing 6ml of Heptane and 6ml of 0.4% Paraformaldehyde.
Shake on orbital rotator at 150rpm for 20 minutes.
Upon completion of 20 minute fixation, allow heptane and parformaldehyde to seperate.
Embyos will be trapped at the interface between the two solutions.
Use a glass pipette to remove the bottom (parafomaldehyde) layer be sure to not disrupt the embryos at the interface.
