Saed Ghassan M
Department of Obstetrics and Gynecology & Cell Biology and Anatomy, Wayne State University School of Medicine, Detroit, MI, USA.
Methods Mol Biol. 2008;477:219-28. doi: 10.1007/978-1-60327-517-0_17.
Immunohistochemistry is an important tool that is often used for the diagnosis of several diseases in the pathology laboratory. The quality and sensitivity of immunohistochemical staining is affected by formalin fixation, which results in variable loss of antigenicity, known as a masking effect. While the sensitivity of immunohistochemistry is excellent for certain antigens, other antigens such as COX-1 and COX-2 are difficult to identify, especially in formalin-fixed, paraffin sections. Antigen retrieval is a technique that re-exposes epitopes and allows detection of masked antigens with standard immunohistochemical procedures. One common method involves partial, enzymatic pre-digestion with trypsin or pepsin while other, nonenzymatic procedures or heat-mediated antigen retrieval methods include pressure-cookers, hot plates, or microwave (MW) irradiation of tissue sections in water or a variety of antigen-retrieval solutions. In this chapter, we will describe a technique that provides a more reliable, much simpler approach for the demonstration of cyclooxygenase-1 and cyclooxygenase-2 expression in frozen, vibratome or paraffin sections, and/or cells in cultures.
免疫组织化学是病理实验室常用于多种疾病诊断的重要工具。免疫组织化学染色的质量和敏感性受福尔马林固定影响，这会导致抗原性出现不同程度的丧失，即所谓的封闭效应。虽然免疫组织化学对某些抗原的敏感性极佳，但其他抗原，如COX - 1和COX - 2，难以识别，尤其是在福尔马林固定的石蜡切片中。抗原修复是一种重新暴露表位并允许通过标准免疫组织化学程序检测被封闭抗原的技术。一种常见方法是用胰蛋白酶或胃蛋白酶进行部分酶预消化，而其他非酶程序或热介导的抗原修复方法包括使用高压锅、热板或对水中或各种抗原修复溶液中的组织切片进行微波照射。在本章中，我们将描述一种技术，该技术为在冷冻切片、振动切片或石蜡切片以及/或者培养细胞中显示环氧化酶 - 1和环氧化酶 - 2的表达提供了一种更可靠、简单得多的方法。
