12 Arthropods Other than Insects
125
Materials
- Dissecting microscope
Equipment
- Centrifuge and centrifuge tube
- Petri dishes (40 and 90 mm diameter)
- Dissecting tray (see Chapter 1, section 3 "Tools")
- Pins
- Fine scissors
- Forceps
- Needles
- Pasteur pipette
- Antibiotics (100 IU/ml penicillin G sodium salt and 1 mg/ml strepto- Solutions
mycin sulfate)
- PBSA (Oxoid)
- Leibovitz's L-IS medium (see Appendix 1)
- Ethanol
Chemicals
- Trypsin
- Versene (= EDTA; Gibco)
- Roccal (Winthrop Laboratories)
- Engorged nymphs, whose tissues of the developing adult can be seen Animals
anteriorly inside the nymphal pelt (this stage is reached in 8 to 19
days depending on the temperature)
Procedure
1. Wash nymphs in tap water.
Sterilization
2. Shake the nymphs in two changes of 1: 10 Roccal, and leave them to
soak in a small container for 20 min in 1 : 10 Roccal solution.
3. Rinse the nymphs quickly in ethanol.
4. Wash the nymphs twice in the antibiotic solution.
5. Transfer the nymphs to a petri dish containing PBSA with antibiotics.
1. Fix the nymphs in a PBSA-filled sterile dissecting tray, dorsal side up, Dissection
with pins at their anterior and posterior ends.
2. Cut around the periphery of the body with fine scissors, and remove
the dorsal integument (Fig. 3).
3. Remove the alimentary canal, Malpighian tubules, and rectum using
fine forceps and needles.
4. Remove the tissues consisting mainly of the epidermis, appendages
and the central ganglion of the developing adults from the ventral part
of the nymphal body.
5. Wash these tissues twice in PBSA in a 40-mm petri dish and transfer
the tissues to another dish containing culture medium.
6. Remove the medium and wash the tissues twice in PBSA.
125
Materials
- Dissecting microscope
Equipment
- Centrifuge and centrifuge tube
- Petri dishes (40 and 90 mm diameter)
- Dissecting tray (see Chapter 1, section 3 "Tools")
- Pins
- Fine scissors
- Forceps
- Needles
- Pasteur pipette
- Antibiotics (100 IU/ml penicillin G sodium salt and 1 mg/ml strepto- Solutions
mycin sulfate)
- PBSA (Oxoid)
- Leibovitz's L-IS medium (see Appendix 1)
- Ethanol
Chemicals
- Trypsin
- Versene (= EDTA; Gibco)
- Roccal (Winthrop Laboratories)
- Engorged nymphs, whose tissues of the developing adult can be seen Animals
anteriorly inside the nymphal pelt (this stage is reached in 8 to 19
days depending on the temperature)
Procedure
1. Wash nymphs in tap water.
Sterilization
2. Shake the nymphs in two changes of 1: 10 Roccal, and leave them to
soak in a small container for 20 min in 1 : 10 Roccal solution.
3. Rinse the nymphs quickly in ethanol.
4. Wash the nymphs twice in the antibiotic solution.
5. Transfer the nymphs to a petri dish containing PBSA with antibiotics.
1. Fix the nymphs in a PBSA-filled sterile dissecting tray, dorsal side up, Dissection
with pins at their anterior and posterior ends.
2. Cut around the periphery of the body with fine scissors, and remove
the dorsal integument (Fig. 3).
3. Remove the alimentary canal, Malpighian tubules, and rectum using
fine forceps and needles.
4. Remove the tissues consisting mainly of the epidermis, appendages
and the central ganglion of the developing adults from the ventral part
of the nymphal body.
5. Wash these tissues twice in PBSA in a 40-mm petri dish and transfer
the tissues to another dish containing culture medium.
6. Remove the medium and wash the tissues twice in PBSA.