Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells.
Combining quantitative 2D and 3D image analysis in the serial block face SEM: application to secretory organelles of pancreatic islet cells.
复制标题
DOI：
10.1111/jmi.12276
复制
发表时间：
2015-08
影响因子：
2
通讯作者：
Leapman RD
中科院分区：
文献类型：
--
作者：
Shomorony A;Pfeifer CR;Aronova MA;Zhang G;Cai T;Xu H;Notkins AL;Leapman RD
通过连续块面扫描电子显微镜获得的组织体积超微结构的二维（2D）和三维（3D）分析的组合可以大大缩短从包含数十亿体素的大数据集获得定量信息所需的时间。因此，为了分析特定类型的细胞器的数量或细胞内由细胞器群体包围的总体积，可以使用体视学方法来估计该细胞器的数量密度或体积分数，以分析通过细胞随机选择的2D块面视图，并且通过在连续的块面图像中描绘质膜来将这种估计与3D细胞体积的精确测量联合收割机相结合。这种方法的有效性可以很容易地测试，因为整个三维组织体积是可用的连续块面扫描电子显微镜数据集。我们已经应用这种混合3D/2D技术来确定小鼠胰岛内分泌α和β细胞中分泌颗粒的数量，并且已经能够估计β细胞的总胰岛素含量。
A combination of two‐dimensional (2D) and three‐dimensional (3D) analyses of tissue volume ultrastructure acquired by serial block face scanning electron microscopy can greatly shorten the time required to obtain quantitative information from big data sets that contain many billions of voxels. Thus, to analyse the number of organelles of a specific type, or the total volume enclosed by a population of organelles within a cell, it is possible to estimate the number density or volume fraction of that organelle using a stereological approach to analyse randomly selected 2D block face views through the cells, and to combine such estimates with precise measurement of 3D cell volumes by delineating the plasma membrane in successive block face images. The validity of such an approach can be easily tested since the entire 3D tissue volume is available in the serial block face scanning electron microscopy data set. We have applied this hybrid 3D/2D technique to determine the number of secretory granules in the endocrine α and β cells of mouse pancreatic islets of Langerhans, and have been able to estimate the total insulin content of a β cell.