Castejon O J
Instituto de Investigaciones Biologicas, Facultad de Medicina, Universidad del Zulia, Maracaibo, Venezuela.
Scanning Microsc. 1993 Jun;7(2):725-40.
In the present paper some basic sample preparation techniques for scanning electron microscopy of nervous tissue are described. These basic preparative methods include conventional scanning electron microscopy or slicing technique, ethanol-cryofracturing technique, freeze-fracture method using either liquid nitrogen (slow freezing) or Freon 22 cooled by liquid nigrogen (fast freezing), improved freeze-fracture method with delicate specimen preparation and chromium coating, ultrasonic microdissection, and "creative tearing" technique. Some basic principles, advantages and limitations are critically considered. In addition, some specific applications in neurobiological research are reported. Emphasis is placed upon understanding the sources and nature of artifacts that are likely to be produced in each preparatory step. Examples are given of the results obtained with the different types of nerve tissue preparation, using the cerebellar cortex as a model of the central nervous system. According to the author's experience, the slicing technique is recommended for studying cytoarchitectonic arrangement of gray centers, the ethanol-cryofracturing technique for tracing short intracortical circuits, and the freeze-fracture methods for analysis of nerve cell cytoplasmic and nuclear compartments. An attempt is made to explain results obtained in relation with nerve cell biology.
本文描述了一些用于神经组织扫描电子显微镜检查的基本样品制备技术。这些基本的制备方法包括传统的扫描电子显微镜或切片技术、乙醇冷冻断裂技术、使用液氮（慢速冷冻）或由液氮冷却的氟利昂22（快速冷冻）的冷冻断裂方法、具有精细样品制备和铬涂层的改进冷冻断裂方法、超声显微解剖以及“创造性撕裂”技术。对一些基本原理、优点和局限性进行了批判性考量。此外，还报道了在神经生物学研究中的一些具体应用。重点在于理解每个制备步骤中可能产生的伪像的来源和性质。以小脑皮质作为中枢神经系统的模型，给出了不同类型神经组织制备所获得结果的示例。根据作者的经验，推荐切片技术用于研究灰质中心的细胞构筑排列，乙醇冷冻断裂技术用于追踪皮质内短回路，冷冻断裂方法用于分析神经细胞质和核区室。试图解释与神经细胞生物学相关的所得结果。
