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Informationen zum Autor J. Robin Harris is the editor of Cell Biology Protocols , published by Wiley. John M. Graham is the editor of Cell Biology Protocols , published by Wiley. David Rickwood is the editor of Cell Biology Protocols , published by Wiley. Klappentext As a modern composite scientific discipline, Cell Biology has expanded and moved forward rapidly in recent years. Cell Biologists now require a wide range of techniques, including those of analytical biochemistry and microscopy in all its diverse forms. These are often used alongside the techniques of molecular biology and molecular genetics. This book contains numerous useful protocols, covering light and electron microscopy, cell culture, cell separation, subcellular fractionation, organelle and membrane isolation, and the use of in vitro reassembly systems in Cell Biology. Many of these protocols feature helpful notes and safety information for practical application. The format favours easy use at the bench with space for notes and important safety information. An appendix contains essential analytical information that will prove invaluable to those working on all aspects of cell biology.This book will be of interest to students and more experienced cell biologists, as well as molecular biologists and those working in genomics and proteomics who are looking for cellular techniques to validate their findings within intact cells. Zusammenfassung As a modern composite scientific discipline, Cell Biology has expanded and moved forward rapidly in recent years. Cell Biologists now require a wide range of techniques, including those of analytical biochemistry and microscopy in all its diverse forms. These are often used alongside the techniques of molecular biology and molecular genetics. Inhaltsverzeichnis Preface xi List of Contributors xiii 1 Basic Light Microscopy 1 Minnie O'Farrell Introduction 1 Key components of the compound microscope 2 Techniques of microscopy 6 Protocols 1.1 Setting up the microscope for bright field microscopy 7 1.2 Setting K¿ohler illumination 8 1.3 Focusing procedure 9 1.4 Setting up the microscope for phase contrast microscopy 11 1.5 Setting up the microscope for epifluorescence 14 1.6 Poly-L-lysine coating 18 References 19 2 Basic Electron Microscopy 21 J. Robin Harris Introduction 21 EM methods available 22 Protocols 2.1 Preparation of carbon-formvar, continuous carbon and holey carbon support films 25 2.2 The 'droplet' negative staining procedure (using continuous carbon, formvar-carbon and holey carbon support films) 27 2.3 Immunonegative staining 29 2.4 The negative staining-carbon film technique: cell and organelle cleavage 31 2.5 Preparation of unstained and negatively stained vitrified specimens 33 2.6 Metal shadowing of biological specimens 35 2.7 A routine schedule for tissue processing and resin embedding 37 2.8 Agarose encapsulation for cell and organelle suspensions 39 2.9 Routine staining of thin sections for electron microscopy 40 2.10 Post-embedding indirect immunolabelling of thin sections 42 2.11 Imaging the nuclear matrix and cytoskeleton by embedment-free electron microscopy 44 Jeffrey A. Nickerson and Jean Underwood References 50 3 Cell Culture 51 Anne Wilson and John Graham Cells: isolation and analysis 51 Anne Wilson Mechanical disaggregation of tissue 52 Protocols 3.1 Tissue disaggregation by mechanical mincing or chopping 54 3.2 Tissue disaggregation by warm trypsinization 56 3.3 Cold trypsinization 58 3.4 Disaggregation using collagenase or dispase 60 Anne Wilson
