8 Insecta (Hemiptera)
97
Subprotocol 4
Larval and Imaginal Organs
Certain organs of large larvae or adults can be cultured as explants.
Materials
- Dissecting microscope
Equipment
- Dissecting tray (see chapter 1, section 3 "Tools") with very fine insect
pins
- Maximov slide
- Ophthalmologists' scissors
- Watchmakers' forceps
- Needles
- Pasteur pipette
- Physiological solution
Solutions
- Culture medium
- Larvae large enough to dissect or adults of hemipteran insects
Animals
Procedure
1. Surface sterilize larvae or adults by dipping them into 70% ethanol Sterilization
for 5-10 min.
2. Wash in sterile distilled water.
1. Fill a dissecting tray with physiological solution.
Dissection
2. Remove the wings when adults are used.
3. Fix the insects in the physiological solution with fine pins.
4. Remove the dorsal plates of the abdomens under a dissecting microscope, and expose the inside of the abdomen (Fig. 6).
5. Distinguish each organ. This can be easily done in the physiological
solution.
6. Remove the organ to be cultured.
7. Collect organs from several insects in physiological solution placed
in the depression of a Maximov slide until a sufficient amount of tissue accumulates.
1. Cut the organs into several pieces.
Culture
2. Wash the organ fragments with fresh physiological solution by chang- set-up
ing the solution.
3. Transfer the fragments into a culture vessel with culture medium.