Menco B P
Department of Neurobiology and Physiology, Northwestern University, Evanston, Illinois 60208-3520, USA.
Microsc Res Tech. 1995 Nov 1;32(4):337-56. doi: 10.1002/jemt.1070320408.
Freeze-fracturing and deep-etching are a well-suited set of methods to study membrane and cytoplasmic features. Various approaches are available. Possible variables include tissue preparation, fracturing only or fracturing followed by etching, modes and materials of replication, and various ways of combining freeze-fracturing and/or deep-etching with (immuno)cytochemistry. Freeze-substitution, in particular combined with embedding in methacrylate resins such as the Lowicryls, is becoming rather widely accepted for purposes of ultrastructural (immuno)cytochemistry. Most investigators active in this field agree that this combination yields superior results compared to (immuno)cytochemistry combined with more conventional means of thin section transmission electron microscopy. Yet relatively little information is available on the variations that can occur with different approaches of freeze-substitution immunocytochemistry. This review deals with some of the variations in freeze-fracturing, freeze-etching, and freeze-substitution as applied to olfactory epithelial structures and with the effectiveness of observations obtained by application of the above sets of methods in relating the special morphology of olfactory epithelial cellular structures with those obtained by other approaches. Indeed, the data obtained continue to provide an integral image in which that morphology can be related to the special biochemistry, cell and molecular biology, and electrophysiology of olfactory epithelial structures.
冷冻断裂和深度蚀刻是一套非常适合研究细胞膜和细胞质特征的方法。有多种方法可供选择。可能的变量包括组织制备、仅进行断裂或断裂后进行蚀刻、复制的模式和材料，以及将冷冻断裂和/或深度蚀刻与（免疫）细胞化学相结合的各种方式。冷冻置换，特别是与诸如Lowicryls等甲基丙烯酸酯树脂包埋相结合，在超微结构（免疫）细胞化学方面正变得相当广泛地被接受。该领域的大多数研究人员都认为，与（免疫）细胞化学结合更传统的薄切片透射电子显微镜方法相比，这种组合能产生更好的结果。然而，关于冷冻置换免疫细胞化学不同方法可能出现的变化，相关信息相对较少。本综述探讨应用于嗅觉上皮结构的冷冻断裂、冷冻蚀刻和冷冻置换的一些变化，以及通过应用上述方法组所获得的观察结果在将嗅觉上皮细胞结构的特殊形态与通过其他方法获得的形态相关联方面的有效性。事实上，所获得的数据继续提供一个完整的图像，在其中该形态可以与嗅觉上皮结构的特殊生物化学、细胞和分子生物学以及电生理学相关联。
