Adult fish were anesthetized in 0.4%Tricaine and secured ventral side up in a slotted sponge. Watchmaker forceps were used to remove the surface scales and penetrate the skin, muscle, and pericardial sac. Once exposed, the ventricle was gently pulled at the apex and cut with iridectomy scissors. After surgery, fish were immediately returned to system water. At the specified time points, hearts were removed and fixed in 4% paraformaldehyde overnight at 4°C, washed several times in PBS, equilibrated in 30% sucrose in PBS, and frozen for cryosectioning.
Protocol detail
Cardiac Tissue Extraction from Adult Fish
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Corresponding Organization :
Other organizations : Center of Regenerative Medicine in Barcelona, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine
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Variable analysis
independent variables
- Anesthesia concentration (0.4% Tricaine)
- Time points for heart removal and fixation
- Cardiac tissue morphology and structure
- Orientation of fish (ventral side up in a slotted sponge)
- Surgical technique (use of watchmaker forceps to remove scales and penetrate skin, muscle, and pericardial sac)
- Heart extraction (gently pulling the ventricle at the apex and cutting with iridectomy scissors)
- Post-surgical treatment (immediately returning fish to system water)
- Negative control: Not mentioned
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