3-dimensional digital reconstruction of the murine coronary system for the evaluation of chronic allograft vasculopathy

Abstract

Background: Chronic allograft vasculopathy (CAV) is a major mechanism of graft failure of transplanted organs in humans. Morphometric analysis of coronary arteries enables the quantitation of CAV in mouse models of heart transplantation. However, conventional histological procedures using single 2-dimensional sections limit the accuracy of CAV quantification. The aim of this study is to improve the accuracy of CAV quantification by reconstructing the murine coronary system in 3-dimensions (3D) and using virtual reconstruction and volumetric analysis to precisely assess neointimal thickness. Methods: Mouse tissue samples, native heart and transplanted hearts with chronic allograft vasculopathy, were collected and analyzed. Paraffin embedded samples were serially sectioned, stained and digitized using whole slide digital imaging techniques under normal and ultraviolet lighting. Sophisticated software tools were used to generate and manipulate 3D reconstructions of the major coronary arteries and branches. Results: The 3D reconstruction provides not only accurate measurements but also exact volumetric data of vascular lesions. This virtual coronary arteriography demonstrates that the vasculopathy lesions in this model are localized to the proximal coronary segments. In addition, virtual rotation and volumetric analysis enabled more precise measurements of CAV than single, randomly oriented histologic sections, and offer an improved readout for this important experimental model. Conclusions: We believe 3D reconstruction of 2D histological slides will provide new insights into pathological mechanisms in which structural abnormalities play a role in the development of a disease. The techniques we describe are applicable to the analysis of arteries, veins, bronchioles and similar sized structures in a variety of tissue types and disease model systems. Virtual slides: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/ 3772457541477230.

A Practical Guide to Whole Slide Imaging: A White Paper From the Digital Pathology Association

“…65 As such, 3-D reconstruction of whole slide histologic data is becoming more relevant and has demonstrated value in both tissue visualization and clinical diagnosis. 63,66 Highresolution 3-D histopathologic imagery is especially advantageous in discovering diagnostic patterns because of the improved correlation among imaging modalities such as magnetic resonance imaging, conventional computed tomography, and WSI. [67][68][69][70] The initial step in 3-D reconstruction of virtual slides involves generating serial, ultrathin tissue sections, which are then mounted to glass slides.…”

Digitally reconstructed top‐down view images provide intuitive understanding of the correlation between dermoscopy and histopathology: a good educational tool

“…Subsequently, each whole section was converted into high‐resolution digital data with a high‐speed scanner (Nano Zoomer, Hamamatsu photonics, Hamamatsu, Japan). Scanned data were reconstructed into the 3D image using 3D visualization software (Voloom, Micro Dimensions, Munich, Germany), which enabled us to obtain an arbitrary 2D image and 3D aerial view of the lesions. Melanocytic skin lesions were observed with a dermoscope (DermLite DL3N, 3Gen Inc, San Juan Capistrano, CA, U.S.A.).…”

Optimized SIFTFlow for registration of whole-mount histology to reference optical images

“…In the first step, the objective function [Eq. (2)] is minimized to find the displacement field using Shojaii and Martel: Optimized SIFTFlow for registration of whole-mount histology to reference optical images the initial parameter values from Ref. 33.…”

“…First, they can provide structural information at the microscopic resolution. For example, biologists have used this approach to study the morphology of embryonic development, 1 the coronary system, 2 and mammary glands 3 in murine models, and 3-D histology reconstruction has also been applied to human breast cancer in the study of ductal carcinoma in situ (DCIS). 4 A 3-D reconstruction also has the potential to improve our understanding of heterogeneity in tumors by allowing us to visualize the spatial distribution of different tumor markers.…”