3D Imaging and Quantitative Characterization of Mouse Capillary Coronary Network Architecture

Abstract

Characterization of the cardiac capillary network structure is of critical importance to understand the normal coronary functional properties and coronary microvascular diseases. The aim of our study was to establish an accessible methodology for 3D imaging and 3D processing to quantitatively characterize the capillary coronary network architecture in mice. Experiments were done on C57BL/6J mice. 3D imaging was performed by light sheet microscopy and confocal microscopy on iDISCO+ optical cleared hearts after labelling of the capillary endothelium by lectin injection. 3D images were processed with the open source software ImageJ. Non-visual image segmentation was based of the frequency distribution of the voxel greyscale values, followed by skeletonization and distance mapping. Capillary networks in left and right ventricles and septum were characterized by the volume network density, the fractal dimension, the number of segments and nodes and their ratio, the total network length, and the average length, diameter, and tortuosity of the segments. Scale-dependent parameter values can be impacted by the resolution limit of the 3D imaging technique. The proposed standardized methodology for 3D image processing is easily accessible for a biologist in terms of investment and difficulty level, and allows the quantification of the 3D capillary architecture and its statistical comparison in different conditions.

Computational Identification and 3D Morphological Characterization of Renal Glomeruli in Optically Cleared Murine Kidneys

“…According to a previous study on heart, we established that, as in the brain, the volume shrinkage was minor, around 19%, without deformation [17]. Our results showed an estimated kidney volume shrinkage around 30%, without significant difference in each dimension, suggesting that, compared to the brain and heart, the iDISCO+ method induced a higher shrinkage, though isotropic, in kidneys.…”

Development and diseases of the coronary microvasculature and its communication with the myocardium

“…The fluorescently labeled vasculature can be visualized by confocal microscopy throughout the entire heart after tissue clearing (Kolesov a et al, 2021). Recently faster and high-quality 3D imaging of coronary vasculature is possible using light sheet microscopy (Nicolas & Roux, 2021).…”

“…Coronary microvasculature can be also visualized by plasma membrane staining using lectin WGA that binds to glycoproteins on the cell surface (Figure 1c,d). Recently, the protocol for quantifying the coronary capillary three‐dimension (3D) architecture has been established by using a combination of light sheet with labeling of the capillary endothelium by lectin injection into vessels (Nicolas & Roux, 2021).…”

A three‐dimensional method for morphological analysis and flow velocity estimation in microvasculature on‐a‐chip

“…Few 3D algorithms have been proposed in the literature as applied to in vivo images. [23][24][25] They are based on similar steps, segmentation, and skeletonization, leveraging distance mapping to compute different network features. Furthermore, they take advantage of the third dimension considering a volumetric vascular density.…”

“…In addition, the method can provide the fractal dimension, the number of segments, vessel length, diameter, and tortuosity. 23 These methods have shown that the third dimension can add important information to the analysis, more properly evaluating the 3D system. The circularity of the vessel cross-section represents a clear example of how the 3D information might be used to enhance the characterization of the model.…”