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Simultaneous measurement of RBC velocity, flux, hematocrit and shear rate in vascular networks

Nature Methods volume 7, pages 655–660 (2010)Cite this article

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Abstract

Not all tumor vessels are equal. Tumor-associated vasculature includes immature vessels, regressing vessels, transport vessels undergoing arteriogenesis and peritumor vessels influenced by tumor growth factors. Current techniques for analyzing tumor blood flow do not discriminate between vessel subtypes and only measure average changes from a population of dissimilar vessels. We developed methodologies for simultaneously quantifying blood flow (velocity, flux, hematocrit and shear rate) in extended networks at single-capillary resolution in vivo. Our approach relies on deconvolution of signals produced by labeled red blood cells as they move relative to the scanning laser of a confocal or multiphoton microscope and provides fully resolved three-dimensional flow profiles within vessel networks. Using this methodology, we show that blood velocity profiles are asymmetric near intussusceptive tissue structures in tumors in mice. Furthermore, we show that subpopulations of vessels, classified by functional parameters, exist in and around a tumor and in normal brain tissue.

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Figure 1: Description, validation and application of RTLS and RVFS.
Figure 2: Analysis of cross-sectional flow profiles in tumor vessels undergoing intussusceptive angiogenesis.
Figure 3: Multiparametric phenotypic vessel clustering to compare vessels in tumor, peritumor and contralateral brain regions in a glioma model.

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Acknowledgements

We thank T. Padera for editorial comments and S. Roberge for technical assistance. This work was supported in part by US National Institutes of Health grants R01HL064240 (L.L.M.), R01CA149285 (L.L.M.) and P01CA80124 (R.K.J.) and by a postdoctoral fellowship from the Susan G. Komen Foundation (W.S.K.).

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Authors and Affiliations

  1. Department of Radiation Oncology, Edwin L. Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, USA

    Walid S Kamoun, Sung-Suk Chae, Delphine A Lacorre, Mariela Mitre, Marijn A Gillissen, Dai Fukumura, Rakesh K Jain & Lance L Munn

  2. Thomson Reuters, Corporate Research & Development, New York, New York, USA

    James A Tyrrell

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  1. Walid S Kamoun
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Contributions

W.S.K. conceived and designed the experiment, validated and implemented the technique, collected data and wrote the manuscript. S.-S.C., D.A.L. and M.A.G. implemented the technique and collected data. J.A.T. conceived and designed the experiment and validated the technique. M.M. validated the technique. D.F. conceived and designed the experiment and provided administrative support. R.K.J. provided administrative and financial support and edited the manuscript. L.L.M. conceived and designed the experiment, provided administrative and financial support and wrote the manuscript.

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Correspondence to Lance L Munn.

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Competing interests

R.K.J. has advisory roles at AstraZeneca, Dyax, Enlight, SynDevRx, Millennium (1 time), Genzyme (1 time), Morphosys (1 time), Astellas (1 time) and Regeneron (1 time), and received honoraria for giving a lecture at Pfizer and Genzyme.

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Kamoun, W., Chae, SS., Lacorre, D. et al. Simultaneous measurement of RBC velocity, flux, hematocrit and shear rate in vascular networks. Nat Methods 7, 655–660 (2010). https://doi.org/10.1038/nmeth.1475

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