Home › Cell Biology › Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury
Cell Biology JoVE (Open Access) Citable · DOI

Induction of Myocardial Infarction in Adult Zebrafish Using Cryoinjury

DOI: 10.3791/3666-v
What you'll learn
  • ✓ Perform cryoinjury procedure to induce reproducible myocardial infarction in adult zebrafish
  • ✓ Collect, fix, mount, and section infarcted zebrafish hearts for histological analysis
  • ✓ Identify and characterize massive cell death patterns in cryoinjured ventricular tissue
Protocol

Biopharma Insights Zebrafish represents a valuable model to study the mechanisms of heart regeneration in vertebrates. Here, we present a protocol for induction of a heart infarct in adult zebrafish using cryoinjury. This method results in massive cell death within 20% of the ventricular wall, similar to that observed in mammalian infarcts.

Difficulty
advanced
Total time
~3–4 hours per fish (cryoinjury + fixation + processing); tissue analysis ongoing over days
Model organism
Zebrafish (Danio rerio), adult
Biosafety
BSL-1

Steps

1
Set up cryoinjury equipment and instruments

Prepare and arrange all equipment required for the cryoinjury procedure, including cryogenic tools and anesthesia delivery systems. Ensure proper temperature control and sterile handling conditions.

▶ 02:30
2
Perform cryoinjury on anesthetized zebrafish heart

Induce myocardial infarction by applying cryogenic injury to the ventricular wall of the anesthetized adult zebrafish, creating massive cell death within approximately 20% of ventricular tissue.

▶ 03:18
3
Collect and fix infarcted zebrafish hearts

Rapidly isolate hearts from euthanized fish and immerse in appropriate fixative solution to preserve tissue morphology and prevent degradation.

▶ 05:37
4
Mount fixed hearts for sectioning

Embed fixed zebrafish hearts in mounting medium (e.g., OCT compound or paraffin) and orient them appropriately for subsequent serial sectioning.

▶ 06:33
5
Section mounted hearts into thin slices

Use a cryostat or microtome to cut serial sections of the embedded heart at appropriate thickness for histological examination and imaging.

▶ 07:29
6
Analyze cryoinjury-induced pathology

Examine and document representative sections showing cell death, tissue damage, and infarct morphology to validate the cryoinjury model and characterize regenerative responses.

▶ 08:41
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