Microchemical Analysis of Nervous Tissue
Methods in Life Sciences
- 1st Edition - January 1, 1974
- Imprint: Pergamon
- Author: Neville N. Osborne
- Language: English
- Paperback ISBN:9 7 8 - 1 - 4 8 3 1 - 7 1 5 7 - 9
- eBook ISBN:9 7 8 - 1 - 4 8 3 1 - 8 7 3 8 - 9
Microchemical Analysis of Nervous Tissue focuses on the use of microbiochemical methods in the analysis of nervous tissue, with emphasis on those related to the study of amines,… Read more
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Request a sales quoteMicrochemical Analysis of Nervous Tissue focuses on the use of microbiochemical methods in the analysis of nervous tissue, with emphasis on those related to the study of amines, amino acids, phospholipids, and proteins. Special attention is paid to the choice of biological material and the various procedures used for the isolation by dissection of defined components of the nervous system. Comprised of 10 chapters, this volume begins with an overview of microprocedures used in neurochemistry, followed by a discussion on the importance of choosing the biological material for microanalysis. The isolation of nervous tissue for analysis is then considered, with particular reference to invertebrate neurons; cell components from fresh, "fixed," freeze-dried, and frozen impregnated tissue; and discrete areas of nervous tissue. Subsequent chapters describe some instruments and glassware used in microprocedures, along with the applications of such procedures; general techniques used in microprocedures; microdetermination of phospholipids as well as amines and amino acids as dansyl derivatives; and microelectrophoresis of proteins. This book will be of interest to molecular biologists, microbiologists, physiologists, and neurochemists.
Preface
Acknowledgments
1. General Introduction
2. Choice of Biological Material for Microanalysis
3. Isolation of Nervous Tissue for Analysis
Isolation of Characterized Invertebrate Neurons
Isolation of Cell Components from Fresh Tissue
Isolation of Cell Components from 'fixed' Tissue
Isolation of Cell Components from Freeze-Dried Tissue
Isolation of Cell Components from Frozen Impregnated Tissue
Isolation of Discrete Areas of Nervous Tissue
4. Some Instruments and Glassware Used in Microprocedures
Dissecting Tools
Scalpel
Glass Needles
Nylon and Hair Loops
Hair Points
Pipettes
General Considerations
Their Construction
Using the Pipettes
Microtubes
5. Available Microbiochemical Procedures and Their Application
Nucleic Acids
Methods
Studies
Enzyme Activities
Methods
Studies
Ions
Methods
Studies
Transmitter Substances
Screening of Transmitters
Microchromatography of Amines
Fluorometry of Amines
6. General Techniques Used in Microprocedures
Weight Determination
Disruption of Nervous Tissue, e.g. Homogenization
Dialysis
7. Microdetermination of Amines and Amino Acids as Dansyl Derivatives
Introduction
Theoretical and Practical Considerations
The Reaction
Determination of Optimum pH
Incubation Time
Concentration of Dansyl-Chloride
Deductions
Requirements for the Analysis of Tissue Samples
Apparatus and Instruments
Glassware
Chemicals and Solutions
Sundries
General Procedure
Dansyl-Chloride Solution
Extraction of Amines and Amino Acids
Dansylation
Microchromatography
Identification of Amines and Amino Acids on Microchromatograms
Autoradiography and Measurement of Radioactivity
Quantification
Procedure for Analyzing Neurons
Biological Application of Procedure
Amines and Amino Acids in Characterized Neurons
Determination of Contents of Specific Amino Acids and Tryptamine
Detection of Biochemical Change in Nervous Tissue of Minute Dimensions
Other Studies Involving the Use of Microdansyl Procedures
Determination of Amino Acids in Peptides and Proteins
The Possibility of Using Dansyl-Chloride to Detect Noradrenaline, Adrenaline, Octopamine and Related Compounds in the Microscale
8. Microdetermination of Phospholipids
Introduction
Theoretical and Practical Considerations
Advantages of Microchromatography
Preparation of Microchromatography Plates
Chromatography
Identification of Lipids
Quantification
Remarks about Fluorometry
Autoradiography
Microanalysis of Individual Phospholipids in Nervous Tissue
Extraction
Microchromatography
Identification and Elution
Fluorometric Determination
Requirements for the Analysis of Tissue Samples
Apparatus and Instruments
Glassware
Chemicals and Solutions
Sundries
Application of the Procedure
Characterized Cells
Nervous Tissue
9. Microelectrophoresis of Proteins
Introduction
General Principles of Polyacrylamide Electrophoresis
Microelectrophoresis (Partly Homogeneous Polyacrylamide Separating Gel)
General Principle
Requirements
Procedure
Determination of Rf Values of Proteins
Isolation of Individual Proteins
Autoradiography of Microgels
Microelectrophoresis on Gradient Gels
Preparation of Gradient Gels
Fractionation of Proteins
Fractionation of Proteins Containing SDS
Adaptation of Microelectrophoresis
Ribonucleic Acid Species
Analysis of Different Dehydrogenases
Microanalysis of Brain Proteins
Proteins Related to Learning in Nerve Cells
Analysis of Proteins in Invertebrate Neurons
Developmental Changes
Microisoelectric Focusing (LDH)
Principle of the Microisoelectric Focusing Method for Detecting
Reagents
Extracts
Procedure
Demonstration of LDH Patterns
Alternative Buffer System for Microisoelectric Focusing
10. General Comments
Index
- Edition: 1
- Published: January 1, 1974
- No. of pages (eBook): 240
- Imprint: Pergamon
- Language: English
- Paperback ISBN: 9781483171579
- eBook ISBN: 9781483187389
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