Cruz-Orive L M
J Microsc. 1987 Feb;145(Pt 2):121-42.
A new stereological method is presented whereby the number of particles in a material can be estimated unbiasedly using a few parallel sections from a block of the material, without having to know either section thickness or the distance between sections, and irrespective of particle size, shape and orientation. All that is needed is the final magnification. The method consists of three steps. Firstly, a few particles are sampled with identical probabilities using a disector (Sterio, 1984) of unknown thickness, namely a selector. Secondly, the volume of each sampled particle is unbiasedly estimated by means of point-sampled intercepts (Gundersen & Jensen, 1985) on a few particle transects. Thus, the mean individual particle volume vN in the population is estimated directly without having to refer to any containing volume. Finally, the particle volume fraction VV is estimated on a random section (by point-counting, say) and the numerical density of particles is estimated via the identity NV = VV/vN. Multiplying the estimate of NV with the reference volume we get an estimate of the total number of particles. The selector may be used not only for counting but also for choosing particles with identical probabilities in order to make, say, serial reconstructions of them. The practical details of the counting method are explained with the aid of a worked synthetic example and a real one involving hepatocytic nuclei. The mathematical proofs are given in two appendices.
本文提出了一种新的体视学方法,通过该方法可以使用材料块中的几个平行切片无偏估计材料中颗粒的数量,而无需知道切片厚度或切片之间的距离,且与颗粒的大小、形状和方向无关。唯一需要的是最终放大倍数。该方法包括三个步骤。首先,使用厚度未知的二分体(Sterio,1984),即选择器,以相同的概率对一些颗粒进行采样。其次,通过在一些颗粒横截面上的点采样截距(Gundersen和Jensen,1985)无偏估计每个采样颗粒的体积。因此,无需参考任何包含体积即可直接估计总体中单个颗粒的平均体积vN。最后,在随机切片上估计颗粒体积分数VV(例如通过点计数),并通过NV = VV/vN恒等式估计颗粒的数密度。将NV的估计值乘以参考体积,我们就得到了颗粒总数的估计值。选择器不仅可用于计数,还可用于以相同概率选择颗粒,以便对它们进行例如连续重建。借助一个合成实例和一个涉及肝细胞核的真实实例解释了计数方法的实际细节。两个附录中给出了数学证明。