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发育中的中枢神经系统的图形和三维重建的制备。

The preparation of graphical and three-dimensional reconstructions of the developing central nervous system.

作者信息

Gribnau A A, Lammers C J

出版信息

Acta Morphol Neerl Scand. 1976 Feb;14(1):1-18.

PMID:818882
Abstract

In the development of the central nervous system morphogenesis and histogenesis are two strongly interdependent aspects. Since the histogenesis can only be investigated on microscopical sections the spatial relations of the various structures are lost. Thus the availability of reliable three-dimensional reconstructions is indispensible. The advantage of the bilateral symmetry of the central nervous system is that it creates the possibility to use the median plane as a natural plane of reference in the piling of the reconstructions, provided the material is sectioned perpendicular to that plane. Such a direction of sectioning is the more advisable, since during the histogenesis the outwards migration of the young neuroblasts mainly is radially orientated. The reconstruction method presented here is based upon a comparison of the median sections of two specimens, which are graphically reconstructed from two different series. Both series are sectioned at right angles to the median plane and also in mutually perpendicular direction. Each resulting optimalized graphically reconstructed median section is used as a reference in the correct piling of the three-dimensional reconstruction of the corresponding series, which consists of polystyrene plates. A prerequisite of the method is an exact orientation of the material during embedding, of which the procedure is described. Secondly the two specimens which yield both series must show a high degree of conformity. In embryological terms this means, that the various developmental stages must be accurately defined. The use of criteria which can only be appropriate when referring to the organ under investigation is argumented. In a discussion on the possibilities and limitations of the method a comparison is made with the reconstruction technique which is based upon the use of artificially introduced outer points of reference.

摘要

在中枢神经系统的发育过程中,形态发生和组织发生是两个紧密相互依存的方面。由于组织发生只能在显微镜切片上进行研究,各种结构的空间关系便丧失了。因此,可靠的三维重建至关重要。中枢神经系统双侧对称的优势在于,只要材料垂直于该平面切片,就有可能将正中平面用作重建堆叠时的自然参考平面。这样的切片方向更为可取,因为在组织发生过程中,年轻神经母细胞的向外迁移主要是径向定向的。这里介绍的重建方法基于对两个标本正中切片的比较,这两个标本是从两个不同系列进行图形重建的。两个系列均垂直于正中平面且相互垂直方向切片。每个最终优化的图形重建正中切片都用作相应系列(由聚苯乙烯板组成)三维重建正确堆叠的参考。该方法的一个前提是在包埋过程中材料的精确取向,文中描述了其步骤。其次,产生两个系列的两个标本必须显示出高度的一致性。从胚胎学角度来说,这意味着必须准确界定各个发育阶段。文中论证了仅在参考所研究器官时才适用的标准的使用。在对该方法的可能性和局限性的讨论中,与基于使用人工引入的外部参考点的重建技术进行了比较。

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