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Three-dimensional reconstruction of tumor microvasculature: Simultaneous visualization of multiple components in paraffin-embedded tissue

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Abstract

Three-dimensional (3D) visualization of microscopic structures may provide useful information about the exact 3D configuration, and offers a useful tool to examine the spatial relationship between different components in tissues. A promising field for 3D investigation is the microvascular architecture in normal and pathological tissue, especially because pathological angiogenesis plays a key role in tumor growth and metastasis formation. This paper describes an improved method for 3D reconstruction of microvessels and other microscopic structures in transmitted light microscopy. Serial tissue sections were stained for the endothelial marker CD34 to highlight microvessels and corresponding images were selected and aligned. Alignment of stored images was further improved by automated non-rigid image registration, and automated segmentation of microvessels was performed. Using this technique, 3D reconstructions were produced of the vasculature of the normal brain. Also, to illustrate the complexity of tumor vasculature, 3D reconstructions of two brain tumors were performed: a hemangioblastoma and a glioblastoma multiforme. The possibility of multiple component visualization was shown in a 3D reconstruction of endothelium and pericytes of normal cerebellar cortex and a hemangioblastoma using alternate staining for CD34 and α-smooth muscle actin in serial sections, and of a GBM using immunohistochemical double staining. In conclusion, the described 3D reconstruction procedure provides a promising tool for simultaneous visualization of microscopic structures.

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Abbreviations

GBM:

glioblastoma multiforme

MVP:

microvascular proliferation

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Acknowledgement

This work has been supported by a grant from the Dutch Cancer Society (KUN2003–2975) to Dr Wesseling.

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Authors and Affiliations

  1. Department of Neurology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

    J. M. M. Gijtenbeek

  2. Department of Pathology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

    P. Wesseling, C. Maass, L. Burgers & J. A. W. M. van der Laak

  3. Department of Neurology 326, Radboud University Nijmegen Medical Centre, P.O. Box 9101, Nijmegen, 6500 HB, The Netherlands

    J. M. M. Gijtenbeek

Authors
  1. J. M. M. Gijtenbeek
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  2. P. Wesseling
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  3. C. Maass
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  4. L. Burgers
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  5. J. A. W. M. van der Laak
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Correspondence to J. M. M. Gijtenbeek.

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Gijtenbeek, J.M.M., Wesseling, P., Maass, C. et al. Three-dimensional reconstruction of tumor microvasculature: Simultaneous visualization of multiple components in paraffin-embedded tissue. Angiogenesis 8, 297–305 (2006). https://doi.org/10.1007/s10456-005-9019-4

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  • DOI: https://doi.org/10.1007/s10456-005-9019-4

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