15 Mollusca
165
Subprotocol 3
Mantle Tissues of Bivalves (Bivalvia)
• • Materials
- Glass plate
- Surgical knife
- Scissors
- Forceps
- Membrane filter system
- UVlamp
- Water bath
- Water pump
- Petri dish (6 em diameter)
- T-25 culture flask
- CO 2 incubator
Fig. 3. A cell line derived from
the embryonic tissue of the
snail Biomphalaria glabrata at
the 46th passage. x 200. From
Hansen,E.L., 1976, with permission of Academic Press
Equipment
- Machii's molluscan balanced salt solution (MMBSS; see Appendix 1) Solutions
- Culture medium Pf-35 (see Appendix 1)
- Bivalves
Animals
• • Procedure
1. Keep the bivalves in running seawater, passed through a membrane Sterilization
filter with a 0.45 ~m pore size, under UV illumination for between
overnight and several days.
1. Cut the adductor muscles of the animal (Fig. 4).
Dissection
2. Remove one valve.
3. Cut a strip (50 mm long, 3 mm wide) of nacre-secreting mantle tissue
from the exposed animal.
1. Place the tissue strip in a petri dish containing MMBSS with 2,000 ~g/ Culture
ml penicillin and 500 ~g/ml kanamycin.
set-up