Methods in Cell Biology

Name: Methods in Cell Biology
Brand: Academic Press
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Academic Press

Published on 23. May 1977

489 pages

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978-0-08-085912-5 (ISBN)

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Cover
Contributors to This Volume
Contents
List of Contributors
Preface
Chapter 1. Cell Separations by Counterflow Centrifugation
I. Introduction
II. Principles of Counterflow Centrifugation
Ill. Theory of the Mechanics of Counterflow Centrifugation
IV. Derivation of a Suitable Chamber Shape
V. Application of Counterflow Centrifugation to Cell Separations
References
Chapter 2. Separation of Spermatogenic Cells and Nuclei from Rodent Testes
I. Introduction
II. Principles of Separation
III. Preparation of Cell Suspensions
IV. Preparation of Nuclear Suspensions
V. Velocity Sedimentation
VI. Equilibrium Density Gradient Centrifugation
VII. Effect of Preparation of Suspension on Separation
VIII. Discussion
IX. Applications
X. Conclusions
References
Chapter 3. Separation of Mammalian Spermatids
I. Introduction
II. Separation According to Cell Size
III. Separation According to Buoyant Density
IV. Discussion
References
Chapter 4. Isolation of Skeletal Muscle Nuclei
I. Introduction
II. Concentration of Nuclei in Skeletal Muscle
III. Homogenization
IV. Purification on Sucrose Gradients
V. Stability of Nuclei
VI. Detailed Procedures for Isolating Muscle Nuclei
References
Chapter 5. Isolation of Neuronal Nuclei from Rat Brain Cortex, Rat Cerebellum, and Pigeon Forebrain
I. Introduction
II. Materials and Methods
III. Results and Discussion
IV. Conclusion
References
Chapter 6. Isolation of Metaphase Chromosomes from Synchronized Chinese Hamster Cells
I. Introduction
II. Cell Line, Medium, and General Techniques
III. Optimum Concentration of Colcemid and Duration of Treatment for Accumulating Mitotic Cells
IV. Collection of a Large Highly Purified Mitotic Cell Population by Repeated Treatments with a Combination of Colcemid and Harvesting Techniques
V. Biological and Cytological Effects of Single and Repeated Treatments with Colcemid and of Chilling on Chinese Hamster Cells
VI. Isolation of Metaphase Chromosomes from Mitotic Cell Population
VII. Discussion
References
Chapter 7. Use of Metrizamide for Separation of Chromosomes
I. Introduction
II. General Methods
III. Chromosome Characterization
IV. Chromosome Separation
V. Discussion
References
Chapter 8. Isolation of Interphase Chromatin Structures from Cultured Cells
I. Introduction
II. Isolation and Purification of Chromatin Structures
III. Events during Lysis
IV. Properties of Chromatin
V. Applications
References
Chapter 9. Quantitative Determination of Nonhistone and Histone Proteins in Chromatin
I. Introduction
II. Existing Methods for Quantitative Determination of the Nonhistone Moiety in Nucleoprotein Material
III. Cellogel Electrophoresis for Quantitative Nonhistone Determinations
IV. Examples of the Quantitative Determination of Histones and Nonhistones by Cellogel Electrophoresis
V. Discussion
References
Chapter 10. Solubilization of Chromatin with Heparin and the Isolation of Nuclear Membranes
I. Introduction
II. Isolation of Nuclei
III. Solubilization of Chromatin
IV. Isolation of Nuclear Envelopes
V. Summary
References
Chapter 11. Fractionation of Cell Organelles in Silica Sol Gradients
I. Introduction
II. Use of Silica Sols
III. Fractionation of Organelles
IV. Purity and Functionality of Silica-Purified Organelles
References
Chapter 12. Preparation of Photosynthetically Active Particles from Synchronized Cultures of Unicellular Algae
I. Introduction
II. Organisms, Growth Conditions, Synchronization
III. Particle Preparation
IV. Particle Characterization
V. Photosynthetic Reactions of the Particles
VI. Evaluation of Reproducibility of Particle Preparations and Activity
References
Chapter 13. Rapid Isolation of Nucleoli from Detergent-Purified Nuclei of Tumor and Tissue Culture Cells
I. Introduction
II. Isolation of Nuclei from Various Cells with Detergents
Ill. Isolation of Nucleoli from Detergent-Purified Nuclei
IV. Applications
V. Commentary
References
Chapter 14. Isolation of Plasma Membrane Vesicles from Animal Cells
I. General Introduction
II. Membrane Markers
Ill. Cell Disruption
IV. Centrifugal Fractionation of Membrane Vesicles
V. Aqueous Two-Phase or Liquid-Interface Partition
VI. Electrophoretic Techniques
VII. Concluding Remarks
References
Chapter 15. Rapid Isolation of Nuclear Envelopes from Rat Liver
I. Introduction
II . Isolation of Rat Liver Nuclei
III. Isolation of Nuclear Envelopes from Purified Nuclei
IV. Characterization of the Basic Nuclear Envelope Preparation
References
Chapter 16. Isolation of Plasma Membranes from Cultured Muscle Cells
I. Introduction
II. Cell Culture Conditions
III. Analytical Procedures
IV. Isolation of Plasma Membranes
V. Distribution, Purification, and Recovery of Membrane Markers
VI. Chemical Composition of Plasma Membranes
VII. Comments
References
Chapter 17. Preparation of Plasma Membranes from Amoebae
I. Introduction
II. Culture Methods
III. Plasma Membrane Isolation
IV. Alternative Isolation Procedures
V. Plasma Membrane Enzymes
VI. Macromolecular Composition of Amoeba Plasma Membrane
VII. Turnover of Plasma Membrane during Phagocytosis
References
Chapter 18. Induction of Mammalian Somatic Cell Hybridization by Polyethylene Glycol
I. Introduction
II. Polyethylene Glycol Solutions
III. Hybridization of Cells in Monolayers
IV. Hybridization of Cells in Suspension
References
Chapter 19. Preparation of Microcells
I. Introduction
II. Micronucleate Cells
III. Preparation of Microcells from Micronucleate Cells
IV. Properties of Microcells
V. Use of Microcells in Somatic Cell Genetics
References
Chapter 20. Nuclear Transplantation with Mammalian Cells
I. Introduction
II. Preparation and Characterization of Karyoplasts
III. Preparation and Characterization of Cytoplasts
IV. Nuclear Transplantation
References
Chapter 21. Procedure for Preparation and Characterization of Liver Cells Made Permeable by Treatment with Toluene
I. Introduction
II. Preparation of Liver Cells Made Permeable by Toluene
Ill. Characterization of the Toluene-Treated Liver Cells
IV. Conclusions
References
Chapter 22. Estimation of Intracellular Fluid Volume
I. Introduction
II. Procedure
III. Results and Discussion
References
Chapter 23. Detection of Carbohydrates with Lectin-Peroxidase Conjugates
I . Introduction
II. Methods of Isolation of Lectins and Preparation of Lectin-HRP Conjugates
III. Fixation
IV. Specificity of Binding between Lectin-HRP and Cell Surface or Intracellular Carbohydrate Moieties
V. Topography, Mobility, and Endocytosis of Surface (Plasma Membrane), Lectin-HRP Conjugates
VI. The Use of Lectin-HRP Conjugates for Demonstration of Intracellular Carbohydrate Moieties in Tissues
VII. Comparison of Lectin-Ferritin to Lectin-HRP Conjugates
VIII. Summary
References
Chapter 24. Measurement of the Growth of Cell Monolayers in Situ
I. Introduction
II. Cells Used in Development of the Technique
III. Handling of Cell Monolayers
IV. Enumeration of Cell Nuclei
V. Number of Fields in Which Counts Should Be Made
VI. Use of the Technique for Testing the Suitability of Different Batches of a Component for a Medium
VII. Estimation of Absolute Numbers of Cells
VIII. Estimates of Specific Cell Populations from Nuclei on Reticule Points Alone
IX. General Discussion
References
Chapter 25. Peptones as Serum Substitutes for Mammalian Cells in Culture
I. Introduction
II. Peptones as Serum Substitutes
III. Cell Growth in Peptonecontaining Medium
IV. Limitations and Prospects
References
Chapter 26. In Situ Fixation and Embedding for Electron Microscopy
I. Introduction
II. Cells on Unsectionable Substrates (Flat-Faced Embedding Technique)
III. Cells on Sectionable Substrates
References
Chapter 27. Genetic and Cell Cycle Analysis of a Smut Fungus ( Ustilago violacea)
I. Introduction
II. Biology and Culture
III. Cell Biology
IV. Genetic Methods
V. Cell Cycle
VI. Mutant Cell Cycle Controls
References
Index
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Methods in Cell Biology

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