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Cell Biology JoVE (Open Access) Citable · DOI

Multiparametric Optical Mapping of the Langendorff-perfused Rabbit Heart

DOI: 10.3791/3160-v
What you'll learn
  • ✓ Harvest and cannulate rabbit hearts for Langendorff perfusion
  • ✓ Conduct optical voltage and calcium mapping using panoramic imaging
  • ✓ Acquire and interpret multiparametric cardiac electrophysiology data
Protocol

This article describes the basic procedures for conducting optical mapping experiments in the Langendorff-perfused rabbit heart using the panoramic imaging system, and the dual (voltage and calcium) imaging modality.

Difficulty
advanced
Total time
~3–4 hours per heart (surgical preparation, perfusion setup, imaging acquisition)
Model organism
Rabbit (New Zealand White or similar)
Biosafety
BSL-1

Steps

1
Harvest and perfuse rabbit heart via Langendorff

Excise rabbit heart following anesthesia and prepare aorta for retrograde aortic perfusion with oxygenated Tyrode solution. Establish stabilized perfusion baseline prior to optical mapping.

▶ 02:29
2
Set up panoramic optical mapping system

Mount perfused heart in imaging chamber, apply voltage-sensitive dye, position camera and illumination optics, and calibrate panoramic imaging system to acquire whole-heart electrical activity maps.

▶ 03:33
3
Conduct dual voltage and calcium imaging

Load calcium indicator, configure dual-modality detection optics, and simultaneously record voltage and intracellular calcium transients across the heart surface during paced or spontaneous activity.

▶ 05:39
4
Analyze and interpret optical mapping results

Process acquired voltage and calcium signals, generate activation maps and phase maps, and extract quantitative electrophysiological parameters from representative beats and arrhythmia episodes.

▶ 07:29
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