Larsen J O
Institute of Pathology, Aalborg Hospital, Denmark.
J Neurosci Methods. 1998 Nov 1;85(1):107-18. doi: 10.1016/s0165-0270(98)00129-0.
Quantitative morphological data from nerve cross sections are of great value in experimental, developmental, and pathological studies. Efficient unbiased stereological techniques to estimate the total numbers and absolute size distributions of both myelinated and unmyelinated axons from a small sample of axons are described and evaluated. Axon numbers are estimated with the fractionator technique. Axon areas are estimated with the 2D-nucleator and the point counting technique for myelinated and unmyelinated axons, respectively. The axon perimeter is estimated by use of a line grid, and the myelin sheath thickness by direct orthogonal measurements in uniform, random locations. The empirically evaluated variance of the number estimates of myelinated axons was approximately approximately 0.05 when approximately 200 axons were counted. The empirical variance of area estimates obtained with a four-way 2D-nucleator was approximately 0.10 and contributed less than 0.5% to the observed total variance of axon areas within an animal. A light microscope modified for stereology with computer assisted stereological test systems and a motorised stage facilitate the acquisition of data for the myelinated axons, which is of primary interest in most studies. The unmyelinated axons are evaluated at the more time consuming electron microscopical level.
来自神经横切面的定量形态学数据在实验、发育和病理学研究中具有重要价值。本文描述并评估了高效、无偏倚的体视学技术，用于从小样本轴突中估计有髓和无髓轴突的总数及绝对大小分布。轴突数量采用分式抽样技术进行估计。有髓和无髓轴突的轴突面积分别采用二维核计数法和点计数法进行估计。轴突周长通过线栅法估计，髓鞘厚度通过在均匀、随机位置进行直接正交测量来确定。当计数约200个轴突时，经经验评估的有髓轴突数量估计值的方差约为0.05。使用四路二维核计数法获得的面积估计值的经验方差约为0.10，对动物体内轴突面积的总观察方差贡献小于0.5%。配备计算机辅助体视学测试系统和电动载物台的用于体视学的改良光学显微镜有助于获取有髓轴突的数据，这在大多数研究中是主要关注点。无髓轴突则在耗时更长的电子显微镜水平进行评估。
