Rapid and improved method for windowing eggs accessing the stage X chicken embryo

The chick stage X blastoderm is routinely accessed through a small window in a freshly laid egg. However, windowing severely compromises embryo survival with hatch rates as low as a few percent. We previously reported a simple modification to the standard method that reduced introduction of air into the sealed egg and improved the hatchability to 32%. Here, we describe an even simpler and more rapid method for sealing a windowed egg using hot glue or paraffin in which the hatch rate increased to an average of 63% of the unwindowed control eggs. The primary reason for low hatchability can be attributed to air trapped within the egg during windowing and/or leakage during incubation, as shown by increased lethality by artificially introducing air into windowed and sealed eggs. Although the hatch rate was considerably improved, air can still enter the egg during incubation and is likely to account for less than 100% hatchability of the sealed eggs. The success of this new windowing method will facilitate high throughput for the production of transgenic birds and find use in developmental biology, toxicity testing, and avian disease research.