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Full Description
This volume describes state-of-the-art protocols that serve as "recipes" for scientists concentrating on fibrosis research. This book is divided into four sections. Part I focuses on animal models of fibrosis and covers topics such as mimicking fibrosis in the lungs, skin, liver and heart, and generating transgenic mouse models. Part II discusses cell culture systems, where the chapters explore cell types important for the development of fibrosis. Part III looks at the purification, quantification, and analysis of the ECM proteins, and Part IV describes computer-assisted methods such as quantifying fibrillar collagen alignment and exploring the nano-surface of collagen with atomic force microscopy (AFM). 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 tips on troubleshooting and avoiding known pitfalls.
Cutting-edge and practical, Fibrosis: Methods and Protocols is a valuable resource aimed at outstanding quality and repeatability of research experiments in the fibrosis field.
Contents
Human Fibrotic Diseases: Current Challenges in Fibrosis Research.- The Bleomycin Model of Pulmonary Fibrosis.- Intra-Dermal Injections of Bleomycin to Model Skin Fibrosis.- Assessing the Effects of Fibrosis on Lung Function by Light Microscopy-Coupled Stereology.- Transplanting Human Skin Grafts onto Nude Mice to Model Skin Scars.- Hypertrophic Scarring in the Rabbit Ear: A Practical Model for Studying Dermal Fibrosis.- Mouse and Rat Models of Induction of Hepatic Fibrosis and Assessment of Portal Hypertension.- Mouse Models of Corneal Scarring.- Modeling Cardiac Fibrosis in Mice: (Myo)Fibroblast Phenotype after Ischemia.- Characterization of Mesenchymal-Fibroblast Cells using the Col1a2 Promoter/Enhancer.- Isolation and Culture of Primary Murine Hepatic Stellate Cells.- Isolation and Culture of Adipose-Derived Stromal Cells from Sub-Cutaneous Fat.- Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting.- Detection of Infiltrating Mast Cells using a Modified Toluidine Blue Staining.- Cell-Populated Collagen Lattice Models.- Traction Force Measurement using Deformable Microposts.- Mechanical Deformation of Cultured Cells with Hydrogels.- Preparation of Decellularized Lung Matrices for Cell Culture and Protein Analysis.- Type I Collagen Purification from Rat Tail Tendons.- Purification of Human Plasma/Cellular Fibronectin and Fibronectin Fragments.- Laser Capture Microdissection of Tissue Sections for High-Throughput RNA Analysis.- Collagen Quantification in Tissue Specimens.- Methods for the Assessment of Active Transforming Growth Factor-β in Cells and Tissues.- Visualizing in Vitro Type I Collagen Fibrillogenesis by Transmission Electron Microscopy.- Histological and Electron Microscope Staining for the Identification of Elastic Fiber Networks.- Method for Picrosirius Red-Polarization Detection of Collagen Fibers in Tissue Sections.- Probing Collagen Organization: Practical Guide for Second Harmonic Generation (SHG) Imaging.- Methods for Quantifying Fibrillar Collagen Alignment.- Exploring the Nano-Surface of Collagenous and Other Fibrotic Tissues with AFM.- Spectral Unmixing Methods and Tools for the Detection and Quantitation of Collagen and Other Macromolecules in Tissue Specimens.- Simple Analysis of Deposited Gene Expression Datasets for the Non-Bioinformatician: How to Use GEO for Fibrosis Research.
