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The process of metastasis formation is hugely complex, as described in the introductory chapter of this book, and this complexity has led us to compile two volumes of methods, from a vastly divergent background that attempts to encompass the whole spectrum of cancer biology. This first volume, Metastasis Research Protocols: Analysis of Cells and Tissues, concentrates on analysis and mapping of molecules produced by cells and tissues and analysis of the molecular biology underlying their expression, whereas the second volume, Metastasis Research Protocols: Cell Behavior In Vitro and In Vivo,focuses sharply on the determination of cell behavior in vitro and in vivo. We have deliberately included chapters describing well-established and familiar te- niques (for example, SDS-PAGE and Western blotting [Chapter 11], and immunocytochemistry [Chapter 2]) in addition to the newer and more speci- ized approaches and specific examples of their application, because-although the methodology is readily available in the published literature and established in many laboratories-we wished these volumes to "stand alone" and to make accessible here the standard techniques that underpin much metastasis research for both the newcomer to the field and the seasoned researcher. Undoubtedly, owing to the complexity of the metastatic cascade and the wealth of research techniques involved in scientific approaches to its unraveling, and despite our best efforts to make these volumes as comprehensive as seems feasible, this is a tall order, and there will inevitably be omissions. For these we apologize.
List of contents
Detection Methods for Cellular Markers of Metastatic Potential.- Histopathological Assessment of Metastasis.- Basic Immunocytochemistry for Light Microscopy.- Multiple Labeling Techniques for Fluorescence Microscopy.- Lectin Histochemistry to Detect Altered Glycosylation in Cells and Tissues.- Immunocytochemical Detection and Characterization of Individual Micrometastatic Tumor Cells.- Understanding Metastasis Through Integrin Expression.- Surrogate Markers of Angiogenesis and Metastasis.- Measurement of Microvessel Density in Primary Tumors.- Assessment of Cellular Proliferation by Calculation of Mitotic Index and by Immunohistochemistry.- Flow Cytometric Measurement of Cell Proliferation.- SDS -PAGE and Western Blotting Techniques.- Assessment of Gelatinases (MMP-2 and MMP-9 by Gelatin Zymography.- Genetic Aspects of Metastasis.- In Situ Hybridization to Localize mRNAs.- Fluorescence itIn Situ Hybridization.- Comparative Genomic Hybridization.- RNA Purification and Quantification Methods.- Northern and Southern Blotting and the Polymerase Chain Reaction to Detect Gene Expression.- Methylation Analysis of CpG Islands.- Detection of MetastaticTumor Cells in Blood by Reverse Transcriptase-Polymerase Chain Reaction.- Differential Display.- Mathematical Modeling of Metastasis.- Predictive Mathematical Modeling in Metastasis.
