Method for treating and processing whole chick embryos for autoradiography, immunocytochemistry and other techniques.

A method is presented for in situ treatment of whole chick embryos with drugs and immunocytochemical and fixative reagents that resembles conditions "in ovo." The chick embryo is placed in a "shell-less" culture system where it is contained by an agar ring allowing for treatment in vivo. The conceptus (embryo+membranes) is then mounted on a microporous membrane and inserted into a filter device connected to a three-way stopcock that permits fluids to be changed using syringes. The embryo is then processed in toto or after embedding and sectioning for light or electron microscopy. The proposed handling system decreases technical artifacts and changes in the topographic microanatomy produced by conventional manipulation of chick embryos. This method is useful also for directly observing and recording changes in the embryo during drug treatments and allows processing with dangerous reagents without their direct contact with the operator. It is simple, inexpensive and requires only minimal technical training.

[1]  R. Rovasio,et al.  Exogenous retinoic acid decreases in vivo and in vitro proliferative activity during the early migratory stage of neural crest cells , 1997, Cell proliferation.

[2]  R. A. Rovasio,et al.  Role of early migratory neural crest cells in developmental anomalies induced by ethanol. , 1995, The International journal of developmental biology.

[3]  M. G. Paglini,et al.  Cell cycle of neural crest cells in the early migratory stage in vivo , 1994 .

[4]  V. Hamburger,et al.  A series of normal stages in the development of the chick embryo. 1951. , 2012, Developmental dynamics : an official publication of the American Association of Anatomists.

[5]  B. Monis,et al.  Carrageenan Induces Anomalies in the Chick Emhryo. A Light Microscopic Study , 1987, Toxicologic pathology.

[6]  J. Thiery,et al.  A cell surface marker for neural crest and placodal cells: further evolution in peripheral and central nervous system. , 1984, Developmental biology.

[7]  R. A. Rovasio,et al.  Teratogenic effect of lambda-carrageenan on the chick embryo. , 1981, Teratology.

[8]  B. Monis,et al.  Lethal and Teratogenic Effects of Lambda—Carrageenan, A Food Additive, On the Development of the Chick Embryo , 1980 .

[9]  James G. Wilson Survey of in Vitro Systems: Their Potential Use in Teratogenicity Screening , 1978 .

[10]  J. Folkman,et al.  A simple procedure for the long-term cultivation of chicken embryos. , 1974, Developmental biology.

[11]  O. Fitzhugh,et al.  THE INJECTION OF CHEMICALS INTO THE YOLK SAC OF FERTILE EGGS PRIOR TO INCUBATION AS A TOXICITY TEST. , 1963, Toxicology and applied pharmacology.