Rydmarker S, Dunn D E, Nilsson P, Lindqvist C
Scan Electron Microsc. 1986(Pt 4):1459-67.
Scanning electron microscopy (SEM) permits a three-dimensional study of the surface morphology of the organ of Corti that is very useful in evaluating the condition of the apical end of the hair cells and the stereocilia. However, some laboratories have experienced problems with curling of the basilar membrane during critical point drying of cochlear specimens prepared for SEM evaluation using the Murakami or osmium thiocarbohydrazide-procedures. This curling of the basilar membrane can obstruct the view of the reticular lamina and the ciliary ends of the hair cells. We have used a dissection method, referred to as the anchor technique, to overcome basilar membrane curling. This technique removes all the structures above the reticular lamina but leaves the basilar membrane attached to the spiral ligament and the lateral bone to which the spiral ligament is anchored. Individual cochlear turns are dissected in this manner and mounted on the same examination stub for SEM evaluation. Maintenance of the lateral attachment of the basilar membrane requires additional dissection time but eliminates the problem of curling during critical point drying. An additional benefit is that mounting the individual turns on the same examination stub facilitates evaluation and photomicroscopy of the surface morphology. The anchor technique has been used successfully on the guinea pig and should be appropriate for most mammalian cochleas.
扫描电子显微镜（SEM）能够对柯蒂氏器官的表面形态进行三维研究，这在评估毛细胞顶端及静纤毛的状况时非常有用。然而，一些实验室在使用村上法或硫代碳酰肼锇法为SEM评估制备耳蜗标本的临界点干燥过程中，遇到了基底膜卷曲的问题。基底膜的这种卷曲会妨碍对网状板和毛细胞纤毛末端的观察。我们采用了一种解剖方法，称为锚定技术，来克服基底膜卷曲的问题。该技术去除了网状板上方的所有结构，但使基底膜附着于螺旋韧带以及螺旋韧带所锚定的外侧骨。以这种方式对单个耳蜗蜗管进行解剖，并安装在同一载物台上进行SEM评估。保持基底膜的外侧附着需要额外的解剖时间，但消除了临界点干燥过程中的卷曲问题。另一个好处是，将单个蜗管安装在同一载物台上便于对表面形态进行评估和显微摄影。锚定技术已在豚鼠身上成功应用，应该适用于大多数哺乳动物的耳蜗。
