Neurological Research Laboratory, Bartholin Institute, Kommune Hospitalet, Copenhagen and Stereological Research Laboratory, University of Aarhus, Denmark.
J Microsc. 1999 Oct;196(1):69-73. doi: 10.1046/j.1365-2818.1999.00555.x.
The local deformation and variations in section thickness are studied in 100-μm thick vibratome sections of well-fixed human brain tissue. During processing, including drying on glass slides, the section thickness is reduced to less than half, but close to the edges there is less shrinkage of the section thickness. Close to both surfaces there is a pronounced reduction in the number of neuronal nucleoli. At the scale of the original section, the upper 15 μm and the lower 10 μm are depleted. The loss is most pronounced at the upper surface, which is unprotected during processing. In the central 70% of the section height, where one would ordinarily use an optical disector for sampling, there is no indication of non-uniform shrinkage. The simplest explanation for the observed loss of nucleoli is that all cells opened by the knife may lose their nuclei across an unprotected section surface. The observations do not generalize to other tissues and other preparation techniques, but illustrate the magnitude of some of the problems for uniform sampling and unbiased estimation in very thick sections. The uniform optical disector sampling of nucleoli in thick sections, as opposed to that of cell nuclei, raises a special problem, which is discussed briefly.
在固定良好的人类脑组织的100μm厚振动切片中研究了局部变形和切片厚度变化。在处理过程中，包括在载玻片上干燥，切片厚度减小到不到原来的一半，但靠近边缘处切片厚度的收缩较小。靠近两个表面处，神经元核仁的数量明显减少。在原始切片的尺度上，上部15μm和下部10μm的区域核仁减少。这种损失在上表面最为明显，因为上表面在处理过程中没有受到保护。在切片高度的中央70%区域，通常会在此处使用光学分割器进行采样，没有迹象表明存在不均匀收缩。对于观察到的核仁损失，最简单的解释是，所有被刀切开的细胞可能会在未受保护的切片表面失去其细胞核。这些观察结果并不适用于其他组织和其他制备技术，但说明了在非常厚的切片中进行均匀采样和无偏估计时一些问题的严重程度。与细胞核的均匀光学分割器采样不同，厚切片中核仁的均匀光学分割器采样引发了一个特殊问题，本文对此进行了简要讨论。
