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Improved method for chick whole-embryo culture using a filter paper carrier
S C Chapman1, J Collignon, G C Schoenwolf
1MRC Centre for Developmental Neurobiology, King's College London, Guy's Hospital, London, United Kingdom. susan.chapman@kcl.ac.uk
Insights
A new Early Chick (EC) culture method simplifies early chick embryo manipulation. This technique improves upon existing methods, enabling easier microsurgery and other procedures for developmental biology research.
Area of Science:
- Developmental Biology
- Embryology
- Experimental Biology
Background:
- Chick whole-embryo culture is crucial for studying early embryonic development.
- Existing methods, like New's method, have limitations in manipulation and orientation.
- Early developmental stages require specialized culture techniques.
Purpose of the Study:
- To introduce a simplified and efficient method for early chick whole-embryo culture.
- To enhance the ability to perform various experimental manipulations on chick embryos.
- To provide an improved alternative to existing chick embryo culture techniques.
Main Methods:
- Utilizes a filter paper carrier to maintain tension on the blastoderm and vitelline membranes.
- Employs an agar-albumen substratum for embryo growth.
- The modified method is termed Early Chick (EC) culture.
Main Results:
- EC culture supports chick embryos from pre-primitive streak stages to stage 10.
- Facilitates microsurgery, grafting, bead implantation, microinjection, and electroporation.
- Allows dorsal-side up or ventral-side up culturing for embryos at stage 8 and older.
Conclusions:
- The EC culture method is a significant improvement for early chick embryo manipulation.
- This technique offers greater flexibility in embryo orientation and experimental procedures.
- EC culture provides a valuable tool for advancing research in developmental biology.
Abstract:
We describe a simple method of chick whole-embryo culture, which uses a filter paper carrier to hold the early blastoderm and vitelline membranes under tension while the embryo grows on a substratum of agar-albumen. This is a quick and efficient means of setting up cultures of chick embryos beginning at pre-primitive streak stages to stage 10 (stages X--XIV, Eyal-Giladi and Kochav [1976] Dev Biol 49:321-337; stages 1--10, Hamburger and Hamilton [1951] J Morphol 88:49--92). This is an improvement on the original method of New, which used a glass ring and watch glass (New [1955] Exp Morphol 3:320--331). Our modification of New's method, which we call EC (Early Chick, pronounced EASY) culture, facilitates several manipulations in early chick embryos, including microsurgery, grafting, bead implantation, microinjection, and electroporation. Using the EC method, embryos at stage 8 and older can be readily cultured either dorsal-side up (in contrast to New's method) or ventral-side up, as desired; embryos younger than stage 8 can be culture only ventral-side up (as with New's method). We also discuss some alternative methods for setting up these cultures.