22 Arthropoda Other than Insecta
271
1. Remove the dorsobranchial carapace by splitting the dorsal seam to- Dissection
ward the rostrum, then pulling each half of the carapace free of the
thorax (Fig. 1).
2. Work the underlying integument free of the old cuticle and place the
free integument in complete medium I.
3. Remove the hepatopancreas and other tissues adhering to the integument.
4. Wash the integument in fresh complete medium I.
1. Cut the integument into 3-5-mm 2 pieces, and transfer five fragments Culture
to a Falcon culture dish.
set-up
2. Orient the tissue so that the cuticle faces upward.
3. Cover each plate with 2 ml complete medium I.
4. Maintain the cultures in a humidified incubator at 25°C for the duration of the experiment.
1. Replace the medium after 24 h with 2 ml complete medium I.
Maintenance
2. Every 2 days thereafter, replace the medium with complete medium
II.
Results
According to Ballard et al. (1993), the cuticular epithelium of cultured
pre-exuvial integument remained viable and retained its columnar morphology over a period of 21 days. During the culture period, the tissue
seemed to deposit some additional exocuticule.
Subprotocol 3
Mantle Tissue of Cirripeds (Crustacea)
Materials
- Scissors
- Forceps
- Membrane filter
- Test tubes (100 x 13 mm)
- Test tube rack
- Oscillator
Equipment
- Filter-sterilized seawater
Solutions
- Culture medium (Eagle's minimal essential medium (MEM), supplemented with 4.5 mM N-acetylglucosamine, 100 IU/ml penicillin,
100 Ilglml streptomycin; pH 7.7)
- Barnacles Balanus amphitrite
Animals
271
1. Remove the dorsobranchial carapace by splitting the dorsal seam to- Dissection
ward the rostrum, then pulling each half of the carapace free of the
thorax (Fig. 1).
2. Work the underlying integument free of the old cuticle and place the
free integument in complete medium I.
3. Remove the hepatopancreas and other tissues adhering to the integument.
4. Wash the integument in fresh complete medium I.
1. Cut the integument into 3-5-mm 2 pieces, and transfer five fragments Culture
to a Falcon culture dish.
set-up
2. Orient the tissue so that the cuticle faces upward.
3. Cover each plate with 2 ml complete medium I.
4. Maintain the cultures in a humidified incubator at 25°C for the duration of the experiment.
1. Replace the medium after 24 h with 2 ml complete medium I.
Maintenance
2. Every 2 days thereafter, replace the medium with complete medium
II.
Results
According to Ballard et al. (1993), the cuticular epithelium of cultured
pre-exuvial integument remained viable and retained its columnar morphology over a period of 21 days. During the culture period, the tissue
seemed to deposit some additional exocuticule.
Subprotocol 3
Mantle Tissue of Cirripeds (Crustacea)
Materials
- Scissors
- Forceps
- Membrane filter
- Test tubes (100 x 13 mm)
- Test tube rack
- Oscillator
Equipment
- Filter-sterilized seawater
Solutions
- Culture medium (Eagle's minimal essential medium (MEM), supplemented with 4.5 mM N-acetylglucosamine, 100 IU/ml penicillin,
100 Ilglml streptomycin; pH 7.7)
- Barnacles Balanus amphitrite
Animals