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VIPAR, a quantitative approach to 3D histopathology applied to lymphatic malformations
René Hägerling, … , Peter S. Mortimer, Friedemann Kiefer
René Hägerling, … , Peter S. Mortimer, Friedemann Kiefer
Published August 17, 2017
Citation Information: JCI Insight. 2017;2(16):e93424. https://doi.org/10.1172/jci.insight.93424.
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Clinical Medicine Dermatology Vascular biology

VIPAR, a quantitative approach to 3D histopathology applied to lymphatic malformations

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Abstract

BACKGROUND. Lack of investigatory and diagnostic tools has been a major contributing factor to the failure to mechanistically understand lymphedema and other lymphatic disorders in order to develop effective drug and surgical therapies. One difficulty has been understanding the true changes in lymph vessel pathology from standard 2D tissue sections. METHODS. VIPAR (volume information-based histopathological analysis by 3D reconstruction and data extraction), a light-sheet microscopy–based approach for the analysis of tissue biopsies, is based on digital reconstruction and visualization of microscopic image stacks. VIPAR allows semiautomated segmentation of the vasculature and subsequent nonbiased extraction of characteristic vessel shape and connectivity parameters. We applied VIPAR to analyze biopsies from healthy lymphedematous and lymphangiomatous skin. RESULTS. Digital 3D reconstruction provided a directly visually interpretable, comprehensive representation of the lymphatic and blood vessels in the analyzed tissue volumes. The most conspicuous features were disrupted lymphatic vessels in lymphedematous skin and a hyperplasia (4.36-fold lymphatic vessel volume increase) in the lymphangiomatous skin. Both abnormalities were detected by the connectivity analysis based on extracted vessel shape and structure data. The quantitative evaluation of extracted data revealed a significant reduction of lymphatic segment length (51.3% and 54.2%) and straightness (89.2% and 83.7%) for lymphedematous and lymphangiomatous skin, respectively. Blood vessel length was significantly increased in the lymphangiomatous sample (239.3%). CONCLUSION. VIPAR is a volume-based tissue reconstruction data extraction and analysis approach that successfully distinguished healthy from lymphedematous and lymphangiomatous skin. Its application is not limited to the vascular systems or skin. FUNDING. Max Planck Society, DFG (SFB 656), and Cells-in-Motion Cluster of Excellence EXC 1003.

Authors

René Hägerling, Dominik Drees, Aaron Scherzinger, Cathrin Dierkes, Silvia Martin-Almedina, Stefan Butz, Kristiana Gordon, Michael Schäfers, Klaus Hinrichs, Pia Ostergaard, Dietmar Vestweber, Tobias Goerge, Sahar Mansour, Xiaoyi Jiang, Peter S. Mortimer, Friedemann Kiefer

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Figure 1

Spatial arrangement of blood and lymphatic vessels in the human dermis visualized by light-sheet–based microscopy.

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Spatial arrangement of blood and lymphatic vessels in the human dermis v...
A whole-mount immunostained human skin biopsy from a healthy control was analyzed using light-sheet microscopy (ultramicroscopy). Shown are projections of 3D computer reconstructions generated using the volume visualization framework Voreen. The depicted antigens are indicated in their respective colors on top of the images. ESAM1 served as a general endothelial marker, PROX1 acted as a transcription factor marker, and podoplanin (PDPN) identified lymphatic endothelial cells. (A, C, E, and G) Visualization of the tissue volume with the papillary dermis located at top and the cutaneous plexus at the bottom. (B, D, F, and H) Digitally rotated view of the same specimen, showing the vessels of the papillary plexus viewed en face through the epidermis. (A and C) Papillary and cutaneous lymphatic and blood vessel plexuses are distinctly visible (red arrow, papillary plexus; green arrow, cutaneous plexus). (B, F, and H) Valves, identified by condensed high PROX1 expression (between opposing white arrows), were regularly detected in connecting lymphatic (precollector) vessels of the papillary plexus. Blind-ending lymphatic capillaries are marked by white asterisks. Scale bars: 100 μm.

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