26 Annelida
291
- Pasteur pipettes
- Membrane filters
- Sipunculus nudus
• • Procedure
1. Dry the animals by blotting them with filter paper.
2. Immerse the animal in 99 % ethanol.
3. Flame the anterior extremity of the animal quickly to singe it lightly.
1. Make a puncture around the anterior part of the animal using a sterile
pipette (Fig. 2).
2. Suck out the blood.
3. Collect the blood from 30 animals, and centrifuge the collected blood
at 150 g for 10 min.
4. Freeze the plasma and store it frozen.
5. Thaw the plasma and centrifuge it again at 150 g for 10 min.
6. Sterilize the supernatant by passing it through a membrane filter with
a pore size < 0.45 Ilm.
Animals
Sterilization
Preparation
of plasma
1. Aseptically collect blood according to the procedures given above, and Blood
isolate the urns (vase-shaped clusters of cells bearing prominent ter- collection
minal ciliated cells; either attached to the inner coelomic wall or detached in the body cavity) and enigmatic vesicles (multicellular immobile tissue in a liquid cavity), which exist in the body fluid, using
hand centrifugation. The components of the blood will separate into
Nephrostome
Nephridium
Constrictor
---ntiJ~im
intestine
i.4b"!t+i*- Nerve cord
Fig. 2. Structure of a Sipunclus
291
- Pasteur pipettes
- Membrane filters
- Sipunculus nudus
• • Procedure
1. Dry the animals by blotting them with filter paper.
2. Immerse the animal in 99 % ethanol.
3. Flame the anterior extremity of the animal quickly to singe it lightly.
1. Make a puncture around the anterior part of the animal using a sterile
pipette (Fig. 2).
2. Suck out the blood.
3. Collect the blood from 30 animals, and centrifuge the collected blood
at 150 g for 10 min.
4. Freeze the plasma and store it frozen.
5. Thaw the plasma and centrifuge it again at 150 g for 10 min.
6. Sterilize the supernatant by passing it through a membrane filter with
a pore size < 0.45 Ilm.
Animals
Sterilization
Preparation
of plasma
1. Aseptically collect blood according to the procedures given above, and Blood
isolate the urns (vase-shaped clusters of cells bearing prominent ter- collection
minal ciliated cells; either attached to the inner coelomic wall or detached in the body cavity) and enigmatic vesicles (multicellular immobile tissue in a liquid cavity), which exist in the body fluid, using
hand centrifugation. The components of the blood will separate into
Nephrostome
Nephridium
Constrictor
---ntiJ~im
intestine
i.4b"!t+i*- Nerve cord
Fig. 2. Structure of a Sipunclus