Geoffrey R. Newman, Jan A. Hobot

Resin Microscopy and On-Section Immunocytochemistry (eBook, PDF)

• Format: PDF

Produktbeschreibung

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.

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Produktdetails

• Verlag: Springer Berlin Heidelberg
• Seitenzahl: 273
• Erscheinungstermin: 6. Dezember 2012
• Englisch
• ISBN-13: 9783642569302
• Artikelnr.: 53086736

Inhaltsangabe

I: Resin Embedding.- 1 The Strategic Approach.- 2 The Resins.- 3 Resin Embedding Protocols for Chemically Fixed Tissue.- 4 Cryotechniques.- 5 Methods for Resin Polymerisation.- 6 Handling Resin Blocks.- II: On-Section Immunolabelling.- 7 Strategies in Immunolabelling.- 8 General Considerations.- 9 Immunolabelling Protocols for Resin Sections.- 10 Resin Embedding and Immunolabelling.- Appendix I Examples of Typical Resin Embedding Regimes for Immunocytochemistry.- I.1 Solid Tissue, Pellets and Agar Blocks - Immersion Fixation.- I.2 Solid Tissue - Light Perfusion Fixation.- I.3 Solid Tissue - Very Light Perfusion Fixation.- I.4 Cryosubstitution.- I.5 Project Planner.- Appendix II List of Suppliers.- EM General.- EM Apparatus.- Electron Microscopes.- Flow Cytometry.- References.

Rezensionen

From the reviews of the second edition:

"This comprehensive and valuable book is a must-have for the immunohistochemical laboratory. It extensively covers the field of onsection immunocytochemistry, including epoxy and acrylic resins, a variety of embedding and immunolabelling protocols ... . It is the only book I know covering LR-White and -Gold, the different Lowicryls, and Unicryl in this comprehensive manner. ... it is a very helpful book and because of its extensive theoretical and practical reference a standard for the beginner as well as those already familiar with immunocytochemistry." (Jens Krieger, Microscopy and Analysis, September, 2002)