Lametschwandtner A, Lametschwandtner U, Weiger T
Department of Experimental Zoology, University of Salzburg, Austria.
Scanning Microsc. 1990 Dec;4(4):889-940; discussion 941.
The present paper states very briefly the main steps leading to the technique of scanning electron microscopy (SEM) of vascular corrosion casts. From the terms presently used (injection method, microcorrosion cast, injection replica, vascular corrosion cast, vascular cast) the use of "vascular corrosion cast" for lymphatic and blood vessels is recommended. Specification and pretreatment (kind, volume, dosage of anticoagulants, vasoactive substances and spasmolytica used) of the animals examined are referenced as they are available from the literature. The recommendation is given to pay more attention to these parameters than done so far. The steps necessary for producing reasonable and suitable vascular corrosion casts are critically described. Special attention is paid to the physical and chemical properties of the casting media and their significance for polymerization, shrinkage, casting quality, corrosion resistance, and thermal and spatial stability. Emphasis is also focused on the advantages of cutting the vascular corrosion casts embedded in an ice block by a band saw, a self constructed multi-blade cutting device or a mini wheel-saw placed in the chamber of a cryomicrotome. From the drying methods presently used freeze-drying is stressed because of minimal specimen damage. To render casts conductive in most cases sputter-coating is sufficient. It is recommended to run the SEM with 5-10 kV since the resolution received still reveals all details the casting media presently can replicate. Further the application of scanning electron microscopy of vascular corrosion casts in fully differentiated normal tissue, in pathologic tissue as well as in developing tissues and organs is stated. Lastly possibilities and conditions are discussed under which SEM of vascular corrosion casts can serve to quantify vascular structures in order to make the technique more than pure descriptive.
本文简要阐述了血管铸型扫描电子显微镜术(SEM)技术的主要步骤。从目前使用的术语(注射法、微腐蚀铸型、注射复制品、血管腐蚀铸型、血管铸型)来看,建议使用“血管腐蚀铸型”来指代淋巴管和血管。文中引用了所检查动物的规格及预处理方法(所用抗凝剂、血管活性物质和解痉剂的种类、体积、剂量),这些内容可从文献中获取。建议比以往更加关注这些参数。文中批判性地描述了制作合理且合适的血管腐蚀铸型所需的步骤。特别关注了铸型介质的物理和化学性质及其对聚合、收缩、铸型质量、耐腐蚀性以及热稳定性和空间稳定性的重要性。还重点强调了用带锯、自制的多刃切割装置或置于冷冻切片机腔内的微型轮锯切割嵌入冰块中的血管腐蚀铸型所具有的优势。从目前使用的干燥方法来看,由于对标本损伤最小,所以强调冷冻干燥。在大多数情况下,溅射镀膜足以使铸型具有导电性。建议在5 - 10 kV的电压下进行扫描电子显微镜观察,因为所获得的分辨率仍能揭示铸型介质目前所能复制的所有细节。此外,还阐述了血管腐蚀铸型扫描电子显微镜术在完全分化的正常组织、病理组织以及发育中的组织和器官中的应用。最后讨论了血管腐蚀铸型扫描电子显微镜术能够用于量化血管结构的可能性和条件,以便使该技术不仅仅是单纯的描述性技术。