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Updated in 6/21/2018 8:54:34 AM      Viewed: 361 times      (Journal Article)
Journal of visualized experiments : JoVE (19) (2008)

Method for Culture of Early Chick Embryos ex vivo (New Culture)

Delphine Psychoyos , Richard Finnell
ABSTRACT
FROM JOVE.COM:

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying the formation and patterning of brain, neural tube, somite and heart primordia. Applications of chick embryo culture include electroporation of DNA or RNA constructs in order to analyze gene function, grafts of growth factor coated beads such as FGFs and BMPs , as well as whole mount in situ hybridization and immunohistochemistry. This video demonstrates the different steps in chick embryo culture; First, the embryo is explanted in saline. Then, the embryo is centered on a glass ring. The membranes surrounding the embryo are lifted along the walls of the ring. The ring is then placed in a culture dish containing a pool of albumine. The culture dish is sealed and placed in a humid chamber, where the embryo is cultured for up to 24 hrs. Finally, the embryo is removed from the ring, fixed and processed for further applications. A troubleshooting guide is also presented.
DOI: 10.3791/903      ISSN: 1940-087X     
Notes
Part 1: Bench set up A humid chamber is prepared by placing Kimwipe/ddH2O in plastic chamber. A Falcon tube to collect albumin, dishes for culture, rings, watchglass and waste disposal are placed on bench. Pyrex dish is filled with 1.4 l saline (see notes [a]). Part 2: Embryo is explanted in saline Eggs are removed from the incubator after 16 hrs (stage 4). The egg is opene by tapping the shell with forceps. Shell pieces are removed. The thin albumin is collected in Falcon tube. The thick albumin is removed with forceps. The embryo is placed in a plastic dish inside the saline dish. The remaining albumin is removed with forceps. Part 3: Embryo is centered on ring The yolk sac is tilted with forceps so that embryo faces upwards. The yolk sac is cut at the level of the equator or below. Using fine forceps, the vitelline membrane is swiftly peeled. The vitelline membrane is oriented so that its granular side (non shinny) faces upwards. Using fine Read more...