Nicolas Nabil, Nicolas Nour, Roux Etienne
INSERM, Biologie Des Maladies Cardiovasculaires, University Bordeaux, U1034, F-33600 Pessac, France.
Sensors (Basel). 2021 Nov 9;21(22):7440. doi: 10.3390/s21227440.
The aim of this study was to establish an accessible methodology for the objective identification and 3D morphological characterization of renal glomeruli in mice. 3D imaging of the renal cortex was performed by light sheet microscopy on iDISCO+ optical cleared kidneys of six C57BL/6J mice after labelling of the capillary endothelium by lectin injection. 3D images were processed with the open source software ImageJ, and statistical analysis done with GraphPad Prism. Non-visual delimitation of the external surface of the glomeruli was ensured by greyscale-based thresholding, the value of which was determined from the statistical analysis of the voxel frequency distribution. Exclusion of false-positive identification was done by successive volume- and shape-based segmentation. Renal glomeruli were characterized by their number, surface area, volume, and compactness. Average data were expressed as mean ± SD. The number of glomeruli was equal to 283 ± 35 per mm of renal tissue, representing 1.78 ± 0.49% of the tissue volume. The surface area, volume and compactness were equal to 20,830 ± 6200 µm², 62,280 ± 14,000 µm and 0.068 ± 0.026, respectively. The proposed standardized methodology allows the identification of the renal glomeruli and their 3D morphological characterization, and is easily accessible for biologists.
本研究的目的是建立一种可用于小鼠肾小球客观识别和三维形态表征的便捷方法。在用凝集素注射标记毛细血管内皮后，通过光片显微镜对6只C57BL/6J小鼠的iDISCO+光学透明肾脏的肾皮质进行三维成像。三维图像用开源软件ImageJ进行处理，统计分析用GraphPad Prism完成。通过基于灰度的阈值化确保肾小球外表面的非视觉界定，其值由体素频率分布的统计分析确定。通过连续的基于体积和形状的分割排除假阳性识别。肾小球的特征在于其数量、表面积、体积和致密性。平均数据表示为平均值±标准差。肾小球数量为每毫米肾组织283±35个，占组织体积的1.78±0.49%。表面积、体积和致密性分别为20,830±6200µm²、62,280±14,000µm和0.068±0.026。所提出的标准化方法能够识别肾小球及其三维形态特征，并且生物学家易于使用。
