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

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 alpha-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.

Correlations between Perfusion MR Imaging Cerebral Blood Volume, Microvessel Quantification, and Clinical Outcome Using Stereotactic Analysis in Recurrent High-Grade Glioma

“…Future studies using 3D techniques to quantify microvascular parameters in the entire tissue specimen might improve correlations. 49 Third, we calculated rCBV by normalizing lesion DSC-MR imaging measures with contralateral normal parenchyma, though the tumor specimen microvascular parameters were not similarly normalized. Fourth, we measured rCBV in vivo before surgery and compared it with ex vivo tissue analysis.…”

Relative Cerebral Blood Volume Values to Differentiate High-Grade Glioma Recurrence from Posttreatment Radiation Effect: Direct Correlation between Image-Guided Tissue Histopathology and Localized Dynamic Susceptibility-Weighted Contrast-Enhanced Perfusion MR Imaging Measurements

“…In contrast, tumor recurrence promotes angiogenesis and microvascular proliferation, helping to sustain tumor growth. [8][9][10][11] DSC thus offers a biomarker that potentially correlates with histopathology and provides valuable diagnostic information. [12][13][14] We hypothesized that clinically useful threshold rCBV values, which accurately distinguish PTRE and tumor growth, could be developed by directly correlating localized rCBV values with spatially accurate histopathology in the posttreatment setting.…”

Prominent Microvascular Proliferation in Clinically Aggressive Neuroblastoma

“…It is widely accepted that malignant solid tumors must acquire a new blood supply for their progressive growth to a clinically relevant size and for metastasis (3,4). In contrast to normal vessels, tumor vasculature tends to be disorganized with vessels that are dilated and tortuous and have uneven diameters and excessive branching (5). Microvascular proliferation (MVP) ranges from slight to extreme degrees.…”