21 Insecta
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Results
According to Kiguchi et al. (1990), approximately 80% of cultured embryos developed, and 89% of these began to feed on the diet. The larval
duration and the final size of larvae attained were similar to normally
reared silkworms. The adults obtained using the artificial rearing method
mated normally, and laid almost the same number of fertilized eggs as
normally reared females did. The eggs laid by the adults derived from
the cultured embryos were diapause eggs. When embryos were cultured
without yolk and serosa, embryonic development could not be completed.
Subprotocol 21
In Vivo Culture of DrosophilQ Imaginal Discs (Diptera)
Hadorn (1964) developed a method to passage imaginal discs of D.
melanogaster continuously in adult female flies over many passages and
for more than 1 year. The technique developed by Hadorn may be called
in vivo tissue culture or organ culture. The following is based on the
method of Hadorn (1964).
Materials
- Dissecting microscope
- Maximov slide
- Fine knife
- Forceps
- Needles
- Syringe
- Glass capillary
- Physiological solution
- Matured larvae and female adults of D. melanogaster
Procedure
Equipment
Solutions
Chemicals
Animals
1. Select a male larva just before pupation.
Sterilization
2. Sterilize the surface of the larva by submersion in 70% ethanol for
5 min.
3. Rinse the larva with sterilized distilled water.
1. Dissect the larva in physiological solution placed in the depression of Dissection
a Maximov slide.
2. Isolate a male genital disc.