6 Insecta (Diptera)
77
Subprotocol 3
Ovaries
• • Materials
- Dissecting microscope
- Dissecting tray (see chapter 1, section 3 "Tools")
- Scissors
- Forceps
- Needles
- Maximov slide
- Petri dish
- Pasteur pipette
- Physiological solution
- Culture media
- Female adult dipteran insects
• • Procedure
Equipment
Solutions
Animals
1. Surface sterilize adults by dipping them in 70% ethanol for 5-10 min. Sterilization
2. Remove the wings.
1. Fix the insect in a physiological solution placed in a dissecting tray.
Dissection
2. Make an incision in the abdomen using scissors or by tearing the abdominal integument with forceps.
3. Pull out the ovaries.
4. Wash the ovaries in physiological solution, and remove other attached
tissues.
1. Transfer the ovaries into culture medium placed on a Maximov slide. Culture
2. Cut the ovaries into several pieces in the culture medium.
set-up
3. Transfer the suspension of ovarial fragments to a culture vessel.
Fig. 4. Part of cell sheet formed by
cells migrating from adult ovary of
the mosquito Culex pipiens
molestus. From Kitamura (1965)
with permission. x 75