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 .
