Morphology Methods

The past several decades have witnessed an impressive array of conceptual and techno logical advances in the biomedical sciences. Much of the progress in this area has developed directly as a result of new morphology-based methods that have permitted the assessment of chemical, enzymatic, immunological, and molecular parameters at the cellular and tissue levels. Additional novel approaches including laser capture microdissection have also emerged for the acquisition of homogeneous cell popula tions for molecular analyses. These methodologies have literally reshaped the approaches to fundamental biological questions and have also had a major impact in the area of diagnostic pathology. Much of the groundwork for the development of morphological methods was estab th lished in the early part of the 19 century by Francois-Vincent Raspail, generally acknowledged as the founder of the science of histochemistry. The earliest work in the field was primarily in the hands of botanists and many of the approaches to the under standing of the chemical composition of cells and tissues involved techniques such as microincineration, which destroyed structural integrity. The development of aniline th dyes in the early 20 century served as a major impetus to studies of the structural rather than chemical composition of tissue. Later in the century, however, the focus returned to the identification of chemical constituents in the context of intact cell and tissue structure.

Includes supplementary material: sn.pub/extras

Klappentext
Molecular and cell morphological methods have increased rapidly in number and versatility in recent years and, with the completion of the human genome project, will come into much greater demand for the study of human genes and diseases. In Morphology Methods: Cell and Molecular Biology Techniques, Ricardo Lloyd, md, phd, provides critical and tutorial surveys of the cutting-edge molecular and cell biologic techniques used today in the analysis of cells and tissues in situ. These molecular-based histologic and morphologic techniques are described in step-by-step detail by hands-on experts, often those who have originated or perfected them. Among the techniques presented are laser capture microdissection, in situ hybridization, FISH, confocal laser scanning microscopy, in situ PCR, and tyramide amplification. Applications include the detection of infectious agents and DNA, the diagnosis of undifferentiated tumors and endocrine lesions, the metaphase analysis of whole blood cultures, and clonality analysis. Valuable notes on potential pitfalls and practical tips designed to ensure success are included with each method.
In its integrated, state-of-the-art approach, Morphology Methods: Cell and Molecular Biology Techniques thoroughly details and applies the rich panoply of significant new cell and molecular biologic technologies to all the major problems confronting morphologic studies and diagnostic pathology today.

Zusammenfassung
The earliest work in the field was primarily in the hands of botanists and many of the approaches to the under­ standing of the chemical composition of cells and tissues involved techniques such as microincineration, which destroyed structural integrity.

Inhalt
1 Introduction to Molecular Methods.- 2 Laser Capture Microdissection: Principles and Applications.- Protocol: H&E Staining.- Protocol: Operating the PIX Cell System.- Protocol: DNA Extraction from Collected Cells.- Protocol: RNA Extraction from Collected Cells.- 3 In Situ Hybridization: Detection of DNA and RNA.- Protocol: In Situ Hybridization.- 4 In Situ Detection of Infectious Agents.- Protocol: Detecting Adenovirus.- Protocol: Epstein-Barr Virus (Eber I/II).- 5 Fluorescent In Situ Hybridization.- Protocol: FISH.- 6 Practical Applications of the FISH Technique.- Protocol: Metaphase Analysis from Whole Blood Culture.- Protocol: Metaphase Analysis from Solid Tumors.- Protocol: Interphase Analysis on Cell ImprintsCytology Specimens.- Protocol: Interphase Analysis of Formalin-Fixed Paraffin-Embedded Tissues.- 7 Tyramide Amplification Methods for In Situ Hybridization.- Protocol: TSA by Rapp et al., 1995.- Protocol: CARD/TSA with Nanogold.- Protocol: Tyramide-enhanced FISH with Peroxidase-labeled Oligonucleotides.- 8 Protocols: Ultrastructural In Situ Hybridization.- 9 Quantitation of In Situ Hybridization Analysis.- 10 Protocols: Confocal Laser Scanning Microscopy.- 11 Protocols: Polymerase Chain Reaction.- 12 Protocols: Detection of Nucleic Acids in Cells and Tissues by In Situ Polymerase Chain Reaction.- 13 Immunohistochemistry: Theory and Practice.- 14 Antigen Retrieval Technique for Immunohistochemistry: Principles, Protocols, and Further Development.- Protocol: Antigen Retrieval (AR) Technique.- 15 Tyramide Amplification in Immunohistochemistry.- Protocol: CARD Immunohistochemistry.- 16 Application of Immunohistochemistry in the Diagnosis of Lymphoid Lesions.- 17 Applications of Immunohistochemistry in the Diagnosis of Undifferentiated Tumors.- 18 Applicationsof Immunohistochemistry in the Diagnosis of Endocrine Lesions.- 19 Ultrastructural Immunohistochemistry.- Protocol: Ultrastructural Immunohistochemistry.- 20 Protocols: Clonality Analysis.