Castano P, Marcucci A, Miani A, Morini M, Veraldi S, Rumio C
University of Milan, Institute of Human Anatomy, Italy.
J Microsc. 1994 Sep;175(Pt 3):229-37. doi: 10.1111/j.1365-2818.1994.tb03489.x.
Central neurons and peripheral nervous structures, e.g. cutaneous free endings, perifollicular nets, Meissners corpuscles and intramuscular fibres, were studied using various impregnation methods. The confocal scanning laser microscopes (CSLMs) used were equipped with different laser sources, in order to evaluate their limitations and advantages with these techniques and to contribute to a better understanding of the general morphology of the nervous system. When staining with silver sections with clouds of tiny silver granules which are beyond the resolution power of the conventional light microscope but which show a high reflectivity with the CSLM are obtained. Golgi-Cox mercuric impregnation, however, provides specimens which are precipitate-free, thus ensuring the reliability of information obtained. It does, however, have the disadvantage of being applicable only to the central nervous system. In all cases it is an advantage for the instrument to be fitted with different lasers (e.g. Ar and He-Ne), so as to optimize the images of samples impregnated with different methods. Notwithstanding the possibility that artefacts may distort the geometry of the sample and reduce the resolution, the images presented in this paper show that with careful selection of optical sectioning distances, the use of a suitable stack of sections and, if necessary, the aid of false electronic colours and of partial or complete rotation, it is possible to achieve a more precise interpretation of the morphology and organization of complex structures, such as those of the nervous system.
使用各种浸染方法对中枢神经元和外周神经结构进行了研究,例如皮肤游离末梢、毛囊周围神经网、迈斯纳小体和肌内纤维。所使用的共聚焦扫描激光显微镜(CSLM)配备了不同的激光源,以便评估这些技术的局限性和优势,并有助于更好地理解神经系统的总体形态。在用银染色时,可获得带有微小银颗粒云的切片,这些颗粒超出了传统光学显微镜的分辨率,但在CSLM下具有高反射率。然而,高尔基-考克斯汞浸染法提供的标本无沉淀,从而确保了所获信息的可靠性。不过,它的缺点是仅适用于中枢神经系统。在所有情况下,仪器配备不同的激光(如氩激光和氦氖激光)是有好处的,以便优化用不同方法浸染的样品的图像。尽管伪像可能会扭曲样品的几何形状并降低分辨率,但本文呈现的图像表明,通过仔细选择光学切片距离、使用合适的切片堆栈,并在必要时借助假电子颜色以及部分或完全旋转,有可能对复杂结构(如神经系统结构)的形态和组织实现更精确的解读。