Sanchez J G, Speare D J, Sims D E, Johnson G J
Department of Pathology and Microbiology, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, Canada.
J Comp Pathol. 1997 Aug;117(2):165-70. doi: 10.1016/s0021-9975(97)80033-3.
Studies on the microanatomy of the mucus-rich biofilm surface of normal or damaged teleost skin tissue have been limited because conventional fixation regimes do not effectively retain mucus during tissue preparation. A non-aqueous fixation method, based on a technique devised to retain airway mucous for ultrastructural study, and consisting of the use of an inert perfluorocarbon solvent with osmium teroxide 1%, was successfully used to prepare skin tissues of healthy juvenile rainbow trout. The skin's mucous coat was examined by transmission electron microscopy and the results were compared with those obtained with tissues prepared by a conventional glutaraldehyde-based method. In samples fixed with glutaraldehyde, the cell-surface structures retained were limited to microridges and a poorly discernible glycocalyx layer. In contrast, those fixed by the non-aqueous method had a more clearly demonstrated glycocalyx layer, and a second fibrillar layer, resembling mucus, which was separated from the glycocalyx layer by an electron-lucent zone.
由于传统固定方法在组织制备过程中不能有效保留黏液，因此对正常或受损硬骨鱼皮肤组织富含黏液的生物膜表面进行微观解剖学研究一直受到限制。一种非水固定方法，基于一种为保留气道黏液用于超微结构研究而设计的技术，由使用含1%四氧化锇的惰性全氟碳溶剂组成，已成功用于制备健康幼年虹鳟鱼的皮肤组织。通过透射电子显微镜检查皮肤的黏液层，并将结果与用传统戊二醛法制备的组织所获得的结果进行比较。在用戊二醛固定的样本中，保留的细胞表面结构仅限于微嵴和难以辨认的糖萼层。相比之下，用非水方法固定的样本有更清晰显示的糖萼层，以及第二个类似于黏液的纤维层，该纤维层通过一个电子透明区与糖萼层分离。
