Chapter 22
Arthropoda Other Than Insecta
Subprotocol 1
Integument of Crayfish (Crustacea)
Materials
- Sharp knife
- Scissors
- Forceps
- Scalpel
- Glass vials
- Pasteur pipettes
PROTOCOL
Equipment
- van Harreveld's saline (see Appendix 1) containing streptomycin 1 mgl Sol utions
ml, penicillin 1,000 IU/ml, and fungizone 2.5 jJ.g/ml
- Crayfish integument medium (see Appendix 1)
- Crayfish Astacus leptodactylus
Animals
- Parafilm (American National Can)
Miscellaneous
Procedure
1. Surface sterilize the crayfish with 70% ethanol.
1. Cut the integument of the cephalothorax.
2. Isolate the carapace with the adherent integument, and wash it several times for short periods in van Harreveld's saline containing antibiotics.
1. Carefully insert a piece of Parafilm between the carapace and the hypodermis, and cut pieces, 3 or 4 mm in diameter from the carapacehypodermis with inserted Parafilm, using a sharp knife.
2. Separate the hypodermis from the carapace and the parafilm in van
Harreveld's saline.
1. Wash the tissue two or three times in saline and transfer the tissue to
a small glass vial (5 ml capacity) containing 0.5 ml crayfish integument medium.
Sterilization
Dissection
Separation of
hypodermis
Culture
set-up
Arthropoda Other Than Insecta
Subprotocol 1
Integument of Crayfish (Crustacea)
Materials
- Sharp knife
- Scissors
- Forceps
- Scalpel
- Glass vials
- Pasteur pipettes
PROTOCOL
Equipment
- van Harreveld's saline (see Appendix 1) containing streptomycin 1 mgl Sol utions
ml, penicillin 1,000 IU/ml, and fungizone 2.5 jJ.g/ml
- Crayfish integument medium (see Appendix 1)
- Crayfish Astacus leptodactylus
Animals
- Parafilm (American National Can)
Miscellaneous
Procedure
1. Surface sterilize the crayfish with 70% ethanol.
1. Cut the integument of the cephalothorax.
2. Isolate the carapace with the adherent integument, and wash it several times for short periods in van Harreveld's saline containing antibiotics.
1. Carefully insert a piece of Parafilm between the carapace and the hypodermis, and cut pieces, 3 or 4 mm in diameter from the carapacehypodermis with inserted Parafilm, using a sharp knife.
2. Separate the hypodermis from the carapace and the parafilm in van
Harreveld's saline.
1. Wash the tissue two or three times in saline and transfer the tissue to
a small glass vial (5 ml capacity) containing 0.5 ml crayfish integument medium.
Sterilization
Dissection
Separation of
hypodermis
Culture
set-up