Chapter 21
PROTOCOL
Insecta
Subprotocol 1
Imaginal Discs of Caterpillars (Lepidoptera)
Materials
- Dissecting microscope and dissecting tray (see chapter 1, section 3 Equipment
"Tools")
- Scissors
- Forceps
- Needles
- Pasteur pipettes
- Maximov slides
- Culture vessels
- Physiological saline
Solutions
- Culture medium (containing antibiotics; for example, penicillin
100 IV/ml, streptomycin 100 flg/ml, and fungizone 0.25 mg/ml)
- Matured larvae
Animals
Procedure
1. Surface sterilize larvae by submersion in 70% ethanol for 20 min.
Sterilization
2. Wash the larvae in sterilized distilled water.
1. Fix the larvae in a dissecting tray that contains sterile physiological Dissection
saline.
2. Cut meso thoracic epidermal patches from the larvae.
3. Transfer the patches into fresh physiological saline in the depression
of a Maximov slide.
4. Isolate the wing discs by severing the epidermal connections and cutting away any tracheae or fat bodies.
1. Rinse the discs in fresh physiological saline.
Culture
2. Transfer the discs into culture vessels with culture medium, and keep set-up
them at 25°-27°C.