21 Insecta
257
- Pasteur pipettes
- Culture vessels
- Oxygen-supplying system
- Physiological solution
Solutions
- Culture medium (Leloup's medium 1; see Appendix 1)
- Adults of the blowfly Calliphora erythrocephala
Animals
Procedure
1. Surface sterilize the flies by submersion in 70% ethanol for 10 min.
Sterilization
2. Wash the flies with sterilized distilled water.
1. Dissect out the proventriculus in physiological solution. Leave 2 mm Dissection
of the crop duct and 1-2 mm of the midgut on the proventriculus.
2. Rinse the excised proventriculus several times in fresh physiological
solution.
1. Prepare eight to 10 proventriculi, and pool them in a culture vessel Culture
with 20-25 ml culture medium.
set-up
2. During culture, continuously saturate the medium with oxygen.
Results
According to Becker et al. (1975, 1976), the explants secreted peritrophic
membrane from the beginning of culture. This secretion continued nearly
constantly for 10 h, and then decreased. Secretion had stopped by 35 h.
Subprotocol 1 S
Testes of Various Insects
Since Goldschmidt (1915) cultured the spermatocytes of the cecropia
silkworm Hyalophora cecropia, many investigators have cultured the male
gonads of lepidopteran insects as spermatocysts or whole testes. The
methods of cultivation are similar. The following is the common method
used to culture male gonads of lepidopteran insects.
Materials
- Dissecting microscope and dissecting tray (see chapter 1, section 3 Equipment
"Tools")
- Scissors
- Forceps
- Needles
- Pasteur pipettes