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Quantification of fluorescence angiography in a porcine model

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There is no consensus on how to quantify indocyanine green (ICG) fluorescence angiography. The aim of the present study was to establish and gather validity evidence for a method of quantifying fluorescence angiography, to assess organ perfusion.


Laparotomy was performed on seven pigs, with two regions of interest (ROIs) marked. ICG and neutron-activated microspheres were administered and the stomach was illuminated in the near-infrared range, parallel to continuous recording of fluorescence signal. Tissue samples from the ROIs were sent for quantification of microspheres to calculate the regional blood flow. A software system was developed to assess the fluorescent recordings quantitatively, and each quantitative parameter was compared with the regional blood flow. The parameter with the strongest correlation was then compared with results from an independently developed algorithm, to evaluate reproducibility.


A strong correlation was found between regional blood flow and the slope of the fluorescence curves (ROI I: Pearson r = 0.97, p < 0.001; ROI II: 0.96, p < 0.001) as the normalized slope (ROI I: Pearson r = 0.92, p = 0.004; ROI II: r = 0.96, p = 0.001). There was acceptable correlation of the slope of the curve between two independently developed algorithms (ROI I+II: Pearson r = 0.83, p < 0.001), and good resemblance was found with the Bland-Altman method, with no proportional bias.


Perfusion assessment with quantitative indocyanine green fluorescence angiography is not only feasible but easy to perform with commercially available equipment and readily accessible software.

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Equipment was kindly provided during the experiments by Karl Storz (Karl Storz GmbH and Co. KG, Tüttlingen, Germany). The study was supported by the Danish Cancer Research Foundation. The study sponsors had no role in the design of the study; the collection, analysis, or interpretation of the data; the writing of the manuscript; or the decision to submit the manuscript for publication.

Author contributions

Study conception and design: NN, HSA, RA, RBS, MBSS, MHM, LBS, and MA. Acquisition of data: NN, HSA, RA, RBS, MBSS, and MHM. Analysis and interpretation of data: NN, HSA, RA, RBS, MBSS, MHM, LBS, and MA. Drafting of manuscript: NN. Critical revision of manuscript: NN, HSA, RA, RBS, MBSS, MHM, LBS, and MA

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Correspondence to Nikolaj Nerup.

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The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in animals were in accordance with the ethical standards of the institution, at which the studies were conducted. The study was conducted under the supervision of research veterinarians at the Department of Experimental Medicine, Panum Institute, University of Copenhagen and was approved by the Danish Animal Experiments Inspectorate (No. 2012-15-2934-00186).

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Nerup, N., Andersen, H.S., Ambrus, R. et al. Quantification of fluorescence angiography in a porcine model. Langenbecks Arch Surg 402, 655–662 (2017).

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