E-Book Overview
An overview of the techniques used in modern neuroscience research with the emphasis on showing how different techniques can optimally be combined in the study of problems that arise at some levels of nervous system organization. This is essentially a working tool for the scientist in the laboratory and clinic, providing detailed step-by-step protocols with tips and recommendations. Most chapters and protocols are organized such that they can be used independently, while cross-references between the chapters, a glossary, a list of suppliers and appendices provide further help.
E-Book Content
Contents
Chapter 1 Cytological Staining Methods Robert W. Banks Introduction. . . . . . . . . . . . . . . . . . Subprotocol 1: Fixation, Sectioning and Embedding Subprotocol 2: Ultrastructure . . . . . . . . . . Subprotocol 3: The Golgi Method . . . . . . . . Subprotocol 4: Single-Cell Methods. . . . . . . . References . . . . . . . . . . . . . . . . . .
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Chapter 2 Application of Differential Display and Serial Analysis of Gene Expression in the Nervous System Erno Vreugdenhil, Jeannette de Jong and Nicole Datson Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . Subprotocol 1: Differential Display: Practical Approach . . . . . . . . . . Subprotocol 2: Serial Analysis of Gene Expression (SAGE): Practical Approach Subprotocol 3: Digestion of cDNA with Anchoring Enzyme . . . . . . . . Subprotocol 4: Binding to Magnetic Beads. . . . . . . . . . . . . . . . Subprotocol 5: Addition of Linkers . . . . . . . . . . . . . . . . . . . Subprotocol 6: Tag Release by Digestion with Tagging Enzyme . . . . . . . Subprotocol 7: Blunting Tags . . . . . . . . . . . . . . . . . . . . . Subprotocol 8: Ligation to Ditags . . . . . . . . . . . . . . . . . . . . Subprotocol 9: PCR Amplification of Ditags . . . . . . . . . . . . . . . Subprotocol 10: Ditag Isolation . . . . . . . . . . . . . . . . . . . . Subprotocol 11: Concatemerisation . . . . . . . . . . . . . . . . . . . Subprotocol 12: Cloning Concatemers . . . . . . . . . . . . . . . . . Subprotocol 13: Sequencing . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Chapter 3 Methods Towards Detection of Protein Synthesis in Dendrites and Axons Jan van Minnen and R.E. van Kesteren Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . Subprotocol 1: In Situ Hybridization of Cultured Neurons . . . . . . . . Subprotocol 2: In Situ Hybridization at the Electron Microscopic Level . . Subprotocol 3: Single-Cell Differential mRNA Display . . . . . . . . . Subprotocol 4: Functional Implications of mRNAs in Dendrites and Axons: Metabolic Labeling of Isolated Neurites. . . . . . . . . . . . . . . Subprotocol 5: Intracellular Injection of mRNA . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . .
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Chapter 4 Optical Recording from Individual Neurons in Culture Andrew Bullen and Peter Saggau Introduction . . . . . . . . . . . . . . . . . Outline . . . . . . . . . . . . . . . . . . . . Materials . . . . . . . . . . . . . . . . . . . Procedure . . . . . . . . . . . . . . . . . . . Results . . . . . . . . . . . . . . . . . . . . Troubleshooting . . . . . . . . . . . . . . . . Comments . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . .
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