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TECHNICAL ARTICLE Comparing Vasculogenic Mimicry With Endothelial Cell-lined Vessels: Techniques for 3D Reconstruction and Quantitative Analysis of Tissue Components from Archival Paraffin Blocks Amy Y. Lin, MD,* Zhuming Ai, PhD,w Sang-Chul Lee, PhD,z Peter Bajcsy, PhD,z Jacob Pe’er, MD,y Lu Leach, BS,* Andrew J. Maniotis, PhD,* and Robert Folberg, MD* Key Words: laser scanning confocal microscopy, immunohisto- Abstract: We previously described techniques to generate chemistry, 3D reconstruction, melanoma, vasculogenic mimicry 3-dimensional reconstructions of the tumor microcirculation (Appl Immunohistochem Mol Morphol 2007;15:113–119) using immunofluorescence histochemistry and laser scanning confocal microscopy on serial sections from archival formalin- fixed, paraffin-embedded tissues. By aligning sequential z-stacks in an immersive visualization environment (ImmersaDesk), the lthough pathologists render diagnoses based on need to insert fiduciary markers into tissue was eliminated. In A2-dimensional (2D) histologic sections, considerable this study, we developed methods to stitch overlapping confocal information about the biology of disease processes resides z-series together to extend the surface area of interest well in the appearance of features in interest in 3 dimensions beyond that captured by the confocal microscope objective and (3D). For example, at least 2 different components of developed methods to quantify the distribution of markers of the tumor microcirculation have been
Applied Immunohistochemistry & Molecular Morphology – Wolters Kluwer Health
Published: Mar 1, 2007
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