Technique for Liquid Culture of Early Chick Embryos Suitable for Long Term Live Imaging

This protocol describes the culture of early stage chick embryos in liquid culture with the epiblast side up. We have designed a special culture chamber, in which the embryo is supported by a small layer of heavy silicone oil, effectively sealing of the basal side of the epiblast and forming hypoblast from the surrounding medium. Under these conditions the unincubated embryos develop to the fully extended streak stages (HH4-5) during 20 hrs of culture. These conditions are suitable for high resolution imaging with water immersion objectives in an upright or light sheet microscope.