Skoglund T S, Pascher R, Berthold C H
Institute of Anatomy and Cell Biology, University of Göteborg, Sweden.
J Neurosci Methods. 1996 Dec 28;70(2):201-10. doi: 10.1016/s0165-0270(96)00119-7.
We address three problems concerning the quantitative analysis of nerve cell distribution in the cerebral cortex: (i) preparatory tissue deformation (shrinkage); (ii) difficulties in differentiating between small neurons and astroglia; and (iii) the bias introduced by the counting method. We found that staining with Richardson's solution led to no shrinkage in Vibratome-cut sections of aldehyde-fixed rat brains, but did result in staining of the neurons and left the glial cells unstained. This was in striking contrast to Nissl staining which introduced a linear shrinkage of 20-30% and stained all kinds of cortical cells indiscriminately. A computer-based unbiased counting method was implemented by taking advantage of the stereological procedure referred to as the 'optical disector' (Gundersen, H.J.G. (1986) Stereology of arbitrary particles, J. Microsc., 143: 3-45).
（i）制备组织变形（收缩）；（ii）区分小神经元和星形胶质细胞的困难；以及（iii）计数方法引入的偏差。我们发现，用理查森溶液染色不会导致醛固定大鼠脑振动切片收缩，但确实会使神经元染色而使神经胶质细胞未染色。这与尼氏染色形成鲜明对比，尼氏染色会导致20%-30%的线性收缩，并对各种皮质细胞进行无差别染色。利用被称为“光学分割器”的体视学程序（贡德森，H.J.G.（1986年）任意颗粒的体视学，《显微镜学杂志》，143：3-45）实施了一种基于计算机的无偏差计数方法。
