Ogden G R, Lunny D, Lane E B
Department of Dental Surgery and Periodontology, University of Dundee, UK.
Histochem J. 1996 Dec;28(12):875-82. doi: 10.1007/BF02331391.
In immunohistochemistry, it is well known that the majority of monoclonal antibodies to keratins work best on fresh frozen tissue specimens, yet in clinical practice most biopsies are routinely fixed in formaldehyde. This seriously limits the range of keratins that can be reliably assessed in retrospective studies (particularly where only rare archival material exists) and where subtle changes during tissue differentiation may be important. Antigen retrieval using exposure to microwave radiation is one technique that has been applied successfully to other tumour markers (e.g., p53). However, few papers have used this method when immunolabelling for keratins, in spite of the widespread use of antikeratin antibodies as markers of differentiation. The effect of keratin antigen retrieval using microwave processing was assessed on a range of oral mucosal biopsies, since the oral cavity displays a wide range of keratins. A panel of six well characterized antibodies was chosen: LP34 (Ck1, 5, 6, 18), LH1 (Ck10), LL025 (Ck16), A53 BA2 (Ck19), AE8 (Ck13), and E3 (Ck17). For each specimen, one piece was stored in liquid nitrogen and another piece fixed in formalin. Tissue sections were cut from each and, using the peroxidase avidin biotin technique, keratin expression was recorded for a frozen section, a dewaxed section, and a microwave-heated dewaxed section. Although overall there was a 25% improvement in identification of keratins after microwaving, some antibodies performed better than others. Given that keratins have been shown to be of value in tumour diagnosis, this study suggests that microwave processing of archival material can be a valuable adjunct to such analysis.
在免疫组织化学中，众所周知，大多数针对角蛋白的单克隆抗体在新鲜冷冻组织标本上效果最佳，但在临床实践中，大多数活检标本通常用甲醛固定。这严重限制了在回顾性研究中（特别是在仅有罕见存档材料的情况下）以及在组织分化过程中细微变化可能很重要的情况下能够可靠评估的角蛋白范围。利用微波辐射进行抗原修复是一种已成功应用于其他肿瘤标志物（如p53）的技术。然而，尽管抗角蛋白抗体作为分化标志物被广泛使用，但很少有论文在对角蛋白进行免疫标记时使用这种方法。由于口腔黏膜显示出多种角蛋白，因此对一系列口腔黏膜活检标本评估了利用微波处理进行角蛋白抗原修复的效果。选择了一组六种特征明确的抗体：LP34（细胞角蛋白1、5、6、18）、LH1（细胞角蛋白10）、LL025（细胞角蛋白16）、A53 BA2（细胞角蛋白19）、AE8（细胞角蛋白13）和E3（细胞角蛋白17）。对于每个标本，一块保存在液氮中，另一块用福尔马林固定。从每块标本上切取组织切片，并使用过氧化物酶抗生物素蛋白-生物素技术，记录冷冻切片、脱蜡切片和微波加热脱蜡切片的角蛋白表达情况。尽管总体而言，微波处理后角蛋白的识别率提高了25%，但有些抗体的表现优于其他抗体。鉴于角蛋白已被证明在肿瘤诊断中有价值，本研究表明对存档材料进行微波处理可成为此类分析的有价值辅助手段。
