Home › Analytical Chem › Intra-cardiac Side-Firing Light Catheter for Monitoring Cellular Metabolism using Transmural Absorbance Spectroscopy of Perfused Mammalian Hearts
Analytical Chem JoVE (Open Access) Citable · DOI

Intra-cardiac Side-Firing Light Catheter for Monitoring Cellular Metabolism using Transmural Absorbance Spectroscopy of Perfused Mammalian Hearts

DOI: 10.3791/58992-v
What you'll learn
  • ✓ Set up and position a side-firing optical catheter in perfused mammalian hearts
  • ✓ Perform transmural absorbance spectroscopy to measure oxygen tension and substrate utilization
  • ✓ Analyze spectral data to assess cellular metabolism and membrane potential simultaneously
Protocol

Here we introduce a method for using an intra-ventricle optical catheter in perfused hearts to perform absorbance spectroscopy across the heart wall. The data obtained provides robust information on tissue oxygen tension as well as substrate utilization and membrane potential simultaneously with cardiac performance measures in this ubiquitous preparation.

Difficulty
advanced
Total time
~3–4 hours per heart preparation (including perfusion setup, catheter placement, spectral acquisition, and data analysis)
Model organism
Mammalian heart (perfused isolated preparation)
Biosafety
BSL-1

Steps

1
Position side-firing fiber optic catheter intraventricularly

Insert the side-firing optical catheter into the ventricle of the perfused heart and align it to perform transmural light absorbance across the heart wall.

▶ 00:59
2
Perform transmural absorbance spectroscopy acquisition

Collect spectral data across the heart wall using the positioned catheter to simultaneously measure tissue oxygen tension, substrate utilization, and membrane potential alongside cardiac performance parameters.

▶ 04:41
3
Analyze spectral data and fit difference spectra

Process raw absorbance spectra, perform spectral fitting, and generate difference spectra to quantify metabolic and electrophysiological changes under various experimental conditions.

▶ 06:36
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