ABSTRACT
During the past decade, the use of polyand monoclonal antibodies for in situ cell detection at the light microscopical level has developed into a powerful tool for both research and diagnostic purposes. A large number of technical approaches were developed by many investigators, as reviewed by Polak and Van Noorden [1] and Brandtzaeg [2]. Lately, commercially available immunohistochemical kit systems have also become available for single and double immunoenzyme staining. However, the visualization of multiple antigens in one tissue specimen using different enzyme chromogens is still believed to be restricted to “happy few” with “golden hands.” This belief seems justified by the fact that most multiple staining techniques are published as “applied papers,” describing a particular aim and sometimes including the preparation of enzyme and or hapten conjugates. This approach therefore cannot be regarded as “generally applicable” and “reproducible.” This chapter therefore focuses on double and triple immunoenzyme staining methods that can be fully carried out with commercially available reagents. This chapter offers a guide to the many different multiple staining methods and color com-
binations, which usually frightens off the starter in this field. Furthermore, in Section 2.6, general protocols are provided that are easily adapted to detailed working procedures for many different, multiple staining applications.
