Resin Microscopy and On-Section Immunocytochemistry (Springer Lab Manuals)

Since antibodies tagged with markers have been developed, immunocytochemistry has become the method of choice for identifying tissue substances or for the localisation of nucleic acid in tissue by in situ hybridisation. Resin-embedded tissue is routinely used and new techniques are constantly introduced. Thus, the novice entering these fields has a breathtaking variety of methods open to him. This labmanual covers the embedding of tissue using epoxy resin methods to the more sensitive procedures employing the acrylics. The possibilities and results are discussed so that an understanding of the techniques can be acquired and appropriate choices made. The various resins available and all steps involved in tissue processing, beginning with fixation, as well as the great variety of labelling methods and markers that are commonly used for "on-section" cytochemistry and immunocytochemistry are described, including detailed protocols for the application.

Since antibodies tagged with markers have been developed, immunocytochemistry has become the method of choice for identifying tissue substances or for the localisation of nucleic acid in tissue by in situ hybridisation in molecular biology. Resin-embedded tissue is routinely used and new techniques are constantly introduced. Thus, the novice entering these fields has a breathtaking variety of methods open to him. This laboratory book covers the embedding of tissue using less sensitive epoxy resin methods to the more sensitive procedures employing the acrylics. The possibilities are discussed and results are presented so that an understanding of the techniques can be acquired and appropriate choices made.
In the first part of the book, the background of the various resins available are discussed and information on inexpensive alternative technologies where they exist is provided. The various steps involved in tissue processing, beginning with fixation, are first described in theory, then detailed protocols are presented for their application, including troubleshooting sections. The second part rationalises the great variety of labelling methods that are commonly used for "on-section" cytochemistry and immunocytochemistry, where colloidal gold is currently overwhelmingly the most popular marker. The principles behind the usage of various markers and their limitations and advantages are discussed.