Valletta E A, Bertini M, Sbarbati A
Cystic Fibrosis Center, Ospedale Civile Maggiore, Verona, Italy.
Biotech Histochem. 1996 Sep;71(5):245-50. doi: 10.3109/10520299609117168.
Samples of the respiratory mucosa of children with recurrent respiratory infections suspected of having primary ciliary dyskinesia are routinely fixed with glutaraldehyde before ultrastructural examination. This standard technique, however, may not be optimal for visualizing ciliary components or for preserving several cellular and extracellular structures during dehydration and embedding procedures. In this study, brushes of nasal (28 samples) and/or tracheal (9 samples) mucosa from 32 children with recurrent respiratory tract infections were examined. Twenty-nine samples were fixed with glutaraldehyde supplemented with tannic acid to determine if the ultrastructural analysis of respiratory epithelium and bronchial secretions could be improved. Eight samples were conventionally fixed with glutaraldehyde alone. Lesions of the cellular membrane and damaged cells were easily visualized using tannic acid supplemented fixation. Internal ciliary structures including individual microtubules and dynein arms were also more clearly observed. In addition, the internal structure of microvilli of the respiratory epithelium could be studied and the presence of phospholipid-rich surfactant-like material within nasal and tracheal secretions were visualized after tannic acid supplemented fixation. We suggest that addition of tannic acid during fixation is useful for accurate ultrastructural evaluation of respiratory mucosa in both clinical and experimental situations.
疑似患有原发性纤毛运动障碍的反复呼吸道感染儿童的呼吸道黏膜样本，在进行超微结构检查前通常用戊二醛固定。然而，这种标准技术在脱水和包埋过程中，对于观察纤毛成分或保存多种细胞及细胞外结构可能并非最佳选择。在本研究中，对32名反复呼吸道感染儿童的鼻黏膜（28个样本）和/或气管黏膜（9个样本）刷片进行了检查。29个样本用添加了单宁酸的戊二醛固定，以确定是否能改善呼吸道上皮和支气管分泌物的超微结构分析。8个样本仅用戊二醛常规固定。使用添加单宁酸的固定方法，细胞膜损伤和受损细胞易于观察到。包括单个微管和动力蛋白臂在内的内部纤毛结构也更清晰可见。此外，在添加单宁酸固定后，可研究呼吸道上皮微绒毛的内部结构，并观察到鼻和气管分泌物中富含磷脂的表面活性物质样物质的存在。我们建议，在固定过程中添加单宁酸，对于临床和实验情况下呼吸道黏膜的准确超微结构评估均有用。
