Home › Cell Biology › NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts
Cell Biology JoVE (Open Access) Citable · DOI

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

DOI: 10.3791/4115-v
What you'll learn
  • ✓ Set up biventricular working rabbit heart apparatus
  • ✓ Excise rabbit heart for isolated perfusion
  • ✓ Cannulate heart for biventricular working mode
  • ✓ Acquire pressure, AP, and NADH fluorescence signals
Protocol

Biopharma Insights The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

Difficulty
advanced
Total time
~3-4 hours per heart (including setup)
Model organism
Rabbit (New Zealand White)
Biosafety
BSL-1

Steps

1
Set up perfusion and imaging system

Prepare the Langendorff and working heart perfusion apparatus, including buffers, oxygenation, and fluorescence imaging equipment.

▶ 01:25
2
Excise the rabbit heart

Perform surgical excision of the heart from the anesthetized rabbit, ensuring rapid and careful removal to preserve viability.

▶ 02:44
3
Cannulate for biventricular working mode

Cannulate the aorta and pulmonary artery (or other relevant vessels) to establish biventricular working heart perfusion with controlled preload and afterload.

▶ 03:50
4
Acquire pressure and fluorescence signals

Record left and right ventricular pressures, monophasic action potentials, and NADH fluorescence from the epicardial surface.

▶ 07:21
5
Analyze representative hemodynamic and NADH data

Examine typical traces of pressures, monophasic action potentials, and NADH fluorescence to verify proper heart function and signal quality.

▶ 09:08
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