Histology Protocols

This book provides molecular biologists with the basic histochemical techniques and histologists with the molecular techniques necessary to realize the potential of their resource. Authoritative and cutting-edge, the book covers a wide range of techniques.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 47), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry,histologicalstaining (Chapters10and11),aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 1416) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocols.

Provides an easily accessible reference volume for histological techniques written by leading researchers in the field Details the techniques for investigating the expression of many different types of targets in a variety of tissue and samples from sections and cells to electron microscopy Covers a wide range of techniques from classical histochemistry to detection of mRNA, apoptosis, and hypoxia Step-by-step guide to techniques including extensive notes to aid in troubleshooting techniques Includes supplementary material: sn.pub/extras

Klappentext

With much of what we know about the pathogenesis of disease coming from the systematic and careful study of histological material, histology has proven to be an incredibly valuable area of study. In Histology Protocols, expert researchers in the field contribute detailed methods involving the preparation of tissue, providing an overview of fixation, embedding, and processing, along with the required techniques for the retrieval of RNA from histological sections, followed by sections on routine and specialist histological staining techniques, including immuno-, lectin, and hybridization histochemistry, histological staining, as well as specific methods for the in situ identification of hypoxia and apoptosis, and details on the advances in imaging and image analysis. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Histology Protocols will provide molecular biologists with the basic histochemical techniques and histologists with the molecular techniques necessary to realize the potential of their resource.

Inhalt
Tissue Processing.- Tissue Preparation for Histochemistry: Fixation, Embedding, and Antigen Retrieval for Light Microscopy.- An Optimized RNA Extraction Method from Archival Formalin-Fixed Paraffin-Embedded Tissue.- Laser-Capture Microdissection and Pressure Catapulting for the Analysis of Gene Expression in the Renal Glomerulus.- Staining Techniques.- Immunofluorescence Detection of the Cytoskeleton and Extracellular Matrix in Tissue and Cultured Cells.- Double Immunohistochemistry with Horseradish Peroxidase and Alkaline Phosphatase Detection Systems.- Retrogradely Transported Neuronal Tracers Combined with Immunohistochemistry Using Free-Floating Brain Sections.- High-Pressure Freezing, Chemical Fixation and Freeze-Substitution for Immuno-electron Microscopy.- Lectin Histochemistry for Light and Electron Microscopy.- Duplex In Situ Hybridization in the Study of Gene Co-regulation in the Vertebrate Brain.- Special Stains for Extracellular Matrix.- Active Staining of Mouse Embryos for Magnetic Resonance Microscopy.- Immunohistochemical Detection of Tumour Hypoxia.- In Situ Localization of Apoptosis Using TUNEL.- Imaging Techniques.- Use of Confocal Microscopy for Three-Dimensional Imaging of Neurons in the Spinal Cord.- High-Resolution Confocal Imaging in Tissue.- Software-Based Stacking Techniques to Enhance Depth of Field and Dynamic Range in Digital Photomicrography.- Image Analysis and Quantitative Morphology.