21 Insecta
255
- Forceps
- Needles
- Pasteur pipettes
- Culture vessels
- Carlson's solution
Solutions
- Culture media
- Larvae or pupae of lepidopteran insects
Animals
Procedure
1. Surface sterilize the animals by submersion in 70% ethanol.
Sterilization
2. Wash the animals in sterilized distilled water.
1. Fix the animal on a dissecting tray filled with Carlson's solution.
Dissection
2. Make a longitudinal incision in the animal along the dorsal midline.
3. Excise the silk glands.
1. Remove attached tissues from the silk glands in Carlson's solution.
Culture
2. Transfer the glands into culture vessels with culture medium.
set-up
Results
Mitsuhashi (1965) cultured fragments of silk glands of larvae of the rice
stem borer Chilo suppressalis in CSM-2F medium (see Appendix 1) for
approximately 30 days. During culture, some epithelial cell-like cells migrated from the gland. Chinzei (1975) cultured the anterior parts of silk
glands, taken from spinning-stage larvae to I-day-old pupae from the
silkworm Bombyx mori,in Grace's medium (see Appendix 1). The glands
were maintained in a healthy condition without visible changes for over
2 weeks, and showed degeneration when ecdysteroid was added to the
culture medium. Mano and Mitsuhashi (1981) were able to keep the silk
glands of the greater wax moth Galleria mellonella alive for 1 month,
and observed some cell migration from the epithelium of the glands.
Subprotocol 13
Pheromone Glands of the Indian Meal Moth (Lepidoptera)
Materials
- Dissecting microscope and dissecting tray (see chapter 1, section 3 Equipment
"Tools")
- Scissors
- Forceps
- Needles
255
- Forceps
- Needles
- Pasteur pipettes
- Culture vessels
- Carlson's solution
Solutions
- Culture media
- Larvae or pupae of lepidopteran insects
Animals
Procedure
1. Surface sterilize the animals by submersion in 70% ethanol.
Sterilization
2. Wash the animals in sterilized distilled water.
1. Fix the animal on a dissecting tray filled with Carlson's solution.
Dissection
2. Make a longitudinal incision in the animal along the dorsal midline.
3. Excise the silk glands.
1. Remove attached tissues from the silk glands in Carlson's solution.
Culture
2. Transfer the glands into culture vessels with culture medium.
set-up
Results
Mitsuhashi (1965) cultured fragments of silk glands of larvae of the rice
stem borer Chilo suppressalis in CSM-2F medium (see Appendix 1) for
approximately 30 days. During culture, some epithelial cell-like cells migrated from the gland. Chinzei (1975) cultured the anterior parts of silk
glands, taken from spinning-stage larvae to I-day-old pupae from the
silkworm Bombyx mori,in Grace's medium (see Appendix 1). The glands
were maintained in a healthy condition without visible changes for over
2 weeks, and showed degeneration when ecdysteroid was added to the
culture medium. Mano and Mitsuhashi (1981) were able to keep the silk
glands of the greater wax moth Galleria mellonella alive for 1 month,
and observed some cell migration from the epithelium of the glands.
Subprotocol 13
Pheromone Glands of the Indian Meal Moth (Lepidoptera)
Materials
- Dissecting microscope and dissecting tray (see chapter 1, section 3 Equipment
"Tools")
- Scissors
- Forceps
- Needles