Kubínová L, Janácek J, Guilak F, Opatrný Z
Center of Biomathematics, Institute of Physiology, Academy of Sciences of the Czech Republic, Prague.
Cytometry. 1999 Jun 1;36(2):85-95. doi: 10.1002/(sici)1097-0320(19990601)36:2<85::aid-cyto1>3.0.co;2-3.
The implementation of different methods for estimating the surface area and volume of cells studied by confocal microscopy was developed. The methods were compared from the point of view of their precision, applicability and efficiency.
Interactive stereological methods (spatial grid method, fakir method, Cavalieri principle) as well as automatic digital methods (digital Crofton method, voxel counting, triangulation method, iso-intensity contouring method) were considered. The methods were tested on model geometrical solids and on real volume images consisting of a stack of serial sections encompassing entire tobacco BY-2 cells or cell chains.
It is shown that many of the studied methods are very precise when applied to cells of simple or moderately complex shapes. The automatic digital methods are fast and precise but their applicability is limited by the necessity to segment automatically the object surface and to find an optimal resolution. This limitation is not present in stereological methods which are applied interactively and thus are more time-consuming.
The presented implementations of the fakir method and the Cavalieri principle enable interactive, unbiased and efficient estimation of the cell surface area and volume. The recommended steps for measuring the surface area and/or volume of objects studied by confocal microscopy are described.
开发了用于估计共聚焦显微镜研究的细胞表面积和体积的不同方法。从精度、适用性和效率的角度对这些方法进行了比较。
考虑了交互式体视学方法（空间网格法、法基尔法、卡瓦列里原理）以及自动数字方法（数字克罗夫顿法、体素计数、三角测量法、等强度轮廓法）。这些方法在模型几何实体以及由包含整个烟草BY-2细胞或细胞链的一系列连续切片堆栈组成的真实体积图像上进行了测试。
结果表明，许多研究方法应用于简单或中等复杂形状的细胞时非常精确。自动数字方法快速且精确，但其适用性受到自动分割物体表面和找到最佳分辨率的必要性的限制。体视学方法不存在这种限制，体视学方法是交互式应用的，因此更耗时。
所呈现的法基尔法和卡瓦列里原理的实现能够对细胞表面积和体积进行交互式、无偏且高效的估计。描述了测量共聚焦显微镜研究的物体表面积和/或体积的推荐步骤。
