Arnold W
Department of Otolaryngology, Head and Neck Surgery, Kantonsspital Lucerne, Switzerland.
Acta Otolaryngol. 1988 May-Jun;105(5-6):392-7. doi: 10.3109/00016488809119491.
The application of highly sensitive and specific immunohistochemical methods in routine light microscopy has been rewarded by a great many new observations. The essential underlying antigen-antibody reaction has allowed the recognition and identification of some unknown or unidentified components of the cell. Successful results are best achieved by means of reliable fixation and the use of frozen sections. Frozen sections of the human cochlea are, however, impaired by unavoidable damage to the tissues to be studied. The temporal bone has to be decalcified, resulting in a reduction of the range of suitable methods and an interference with a reliable interpretation of the results. The preservation of the specific antigenicity of the tissues is of paramount importance, allowing the detection of the investigated antigen by the antibody applied. The investigation of different fixatives and various methods of decalcification in tissues possessing some familiar and readily identifiable antigens has confirmed the immunohistochemical suitability of properly fixed and adequately decalcified human temporal bones. The present demonstration deals with some of the principal technical procedures and includes examples of their application for the study of the human inner ear. The goal of our research is the development of reliable immunohistochemical methods of fixation and decalcification to be employed in the study of specimens from patients with Ménière's disease, sudden deafness, progressive loss of hearing, genetic sensorineural syndromes, etc. This will lead to the extension of our knowledge of some unrecognized causes of sensorineural hearing loss.
高灵敏度和特异性免疫组织化学方法在常规光学显微镜检查中的应用带来了许多新的发现。基本的抗原-抗体反应使得能够识别和鉴定细胞中一些未知或未明确的成分。通过可靠的固定和使用冰冻切片能够最好地获得成功结果。然而，人耳蜗的冰冻切片会受到待研究组织不可避免的损伤。颞骨必须进行脱钙，这导致合适方法的范围缩小，并干扰对结果的可靠解读。组织特异性抗原性的保存至关重要，这使得能够通过所应用的抗体检测所研究的抗原。对具有一些熟悉且易于识别抗原的组织中不同固定剂和各种脱钙方法的研究证实，适当固定和充分脱钙的人颞骨在免疫组织化学方面是合适的。本演示涉及一些主要技术程序，并包括它们在人类内耳研究中的应用示例。我们研究的目标是开发可靠的免疫组织化学固定和脱钙方法，用于研究梅尼埃病、突发性耳聋、进行性听力丧失、遗传性感觉神经性综合征等患者的标本。这将扩展我们对一些未被认识的感音神经性听力损失原因的认识。
