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A Guide to Fixation and Preservation of TissuesPublished by: Drug and Market Development Publishing Published: Jul. 1, 2005 - 83 Pages Table of ContentsCHAPTER 1: INTRODUCTION TO TISSUE STORAGE Background Tissues Epithelial Tissue Connective Tissue Muscle Tissue Nerve Tissue Organs CHAPTER 2: TISSUE PRESERVATION Introduction Tissue Fixation and Fixatives Classification, Action, and Applications Fixative Agents Aldehydes Formaldehyde Glutaraldehyde Other Aldehydes Other Cross-Reacting Fixatives Oxidizing Agents Osmium Tetroxide Chromic Acid Protein-denaturing Agents Methyl and Ethyl Alcohol Acetic Acid Fixatives with Unknown Mechanisms Picric Acid Mercuric Chloride Acetone Fixation Mixtures Additives for Fixation Tannic acid Phenol Heavy Metal Solutions Lanthanum Lithium Detergents Microwave (MW) Irradiation Microwave Fixation of Brain MW to Accelerate Fixation Other Fixation Strategies Excluded Volume Fixation Vapor Fixation Freeze-drying Post-fixation Treatment Summary CHAPTER 3: PROCEDURAL FACTORS INVOLVED IN FIXATION Temperature Size of Specimens and Penetration of Fixative Changes in Volume pH and Buffers Osmolarity Concentration of Fixatives Duration of Fixation Fixation of Specific Substances Glycogen Lipids Proteins Mucoids Nucleic Acids and Nucleoproteins Biogenic Amines Enzymes Fixation Artifacts CHAPTER 4: FIXATION PROTOCOLS FOR SPECIFIC SUBSTANCES Glycogen Lipids Elftman’s fixative Reagents Required Method Swank and Davenport’s Fixative Reagents Required Method Mucoids Lillie’s Alcoholic Lead Nitrate Reagents Required Method Cetylpyridinium chloride (C.P.C.) Reagents Required Method Lead Subacetate-ethanol Fixative Reagents Required Method Biogenic Amines Potassium Dichromate-acetate-formaldehyde Fixative Reagents Required Method Recovery of DNA, RNA, and Protein with 70% Ethanol Fixation Technique Low-melt Polyester Embedding Method A: Large Tissue Specimens (several cm size) Method B: Small Tissue Specimens and Biopsies (mm to 2 cm size) Histology Slide Cutting Procedure Paraffin Embedding DNA, RNA, and Protein Analysis Tissue Slide Preparation from Laser Capture Microdissection (LCM) Slide Preparation from Manual Microdissection Sample PCR Amplification of DNA Formalin-fixed, Paraffin-embedded Tissue Summation CHAPTER 5: CRYOPRESERVATION OF TISSUE Hypothermic Storage Vitrification The Cryopreservation Process Cryopreservation Case Study Ovarian Tissue Related Processes Freeze-Drying Supercooling Tissue Storage: Equipments and Supplies CHAPTER 6: SUGGESTED READING APPENDIX FORMULATIONS FOR VARIOUS FIXATIVES Formaldehyde Solutions 10% Neutral Buffer Formalin (4% formaldehyde) Reagents Required Baker’s Formol-calcium (modified) Reagents Required Formol Saline Reagents Required Alcoholic Formaldehyde Reagents Required Paraformaldehyde Reagents Required Method Buffered Formaldehyde-glutaraldehyde 200 mOsm38 Reagents Required Method Alcoholic Fixatives Carnoy’s Fixative Reagents Required Method Methacarn Reagents Required Method Wolman’s Solution Reagents Required Method Acetic Alcohol Formalin Reagents Required Method Picric Acid Fixatives Rossman’s Fluid Reagents Required Method Gendre’s Fluid Reagents Required Method Bouin’s Fluid Reagents Required Method Mercuric Fixatives Buffered Formaldehyde Sublimate Reagents Required Method Zenker’s Fluid Reagents Required Method Helly’s Fluid Reagents Required Method B5 Fixative Reagents Required Method Susa Fluid Reagents Required Method AbstractD&MD’s Guide to the Fixation and Preservation of Tissues provides a step by step, in depth analysis and clarification of tissue fixation and storage strategies, challenges and opportunities. The reader will obtain a clear understanding of the role of tissue preservation in discovery, development, toxicity evaluation and drug target characterization. This Guide presents the latest developments in tissue fixatives and combinations, as well as an overview of recent advances in tissue storage tools and technologies to evaluate human drug targets. Also discussed is how different fixatives work, which macromolecules they target and how they can be utilized in different biological conditions. A detailed analysis of post fixation steps is also covered.Get Full Details About This Report >> |
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