21 Insecta
253
Fig. 6. A pro thoracic gland of
the silkworm Bombyx mori
1. Wash the glands in Carlson's solution, and remove other tissues frag- Culture
ments attached to the glands using needles.
set-up
2. Transfer a single gland into the well of a 96-well test plate with 100 fll
culture medium, or into a sitting drop of 20 fll culture medium placed
in a plastic petri dish.
3. Seal the test plate or petri dish with Para film.
4. Keep the culture in a moist chamber at 25°C .
• • Results
Komiya et al. (1998) tested various media, and consequently found that
MGM-450 without sera was the most suitable for the culture of prothoracic glands. Using this medium, the pro thoracic glands maintained their
ecdysteroid-secreting activity for more than 5 days.
Subprotocol 11
Salivary Glands of Dipteran Insects (Diptera)
• • Materials
- Dissecting microscope
Equipment
- Fine knives
- Forceps
- Needles
- Slide glass
- Maximov slide
- Pasteur pipettes
- Culture vessel (a depression slide glass that is 3.75 x 7.5 cm, and has a
depression of 2.8 cm diameter may be used)
- Culture medium (Grace's medium (see Appendix 1) without Solutions
hemolymph and antibiotics, and mixed 5: 1 v/v with 10% ethanol in
distilled water)