Elsevier S & T

METHODS IN CELL BIOLOGY,VOLUME 9

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Table of contents

Cover
Contributors to This Volumeii
Contentsv
List of Contributorsxi
Prefacexiii
Contents of Previous Volumesxv
Chapter 1. Preparation of Large Quantities of Pure Bovine Lymphocytes and a Monolayer Technique for1
I. Introduction1
II. Preparation of Bovine Lymphocytes3
III. Cultivation of Bovine Small Lymphocytes in a Monolayer6
References10
Chapter 2. Methods to Culture Diploid Fibroblasts on a Large Scale13
I. Introduction13
II. Establishing a Human Fibroblast Culture14
Ill. An Automated Method for Growing Cells on Glass Beads (Perlacell)16
IV. Other Methods for Mass-Scale Culturing of Fibroblasts19
References23
Chapter 3. Partition of Cells in Two-Polymer Aqueous Phases: A Method for Separating Cells and for O25
I. Introduction25
II. Methods35
III. Results40
IV. Concluding Remarks49
References49
Chapter 4. Synchronization of Cell Division in Vivo through the Combined Use of Cytosine Arabinoside51
I. Introduction51
II. Materials and Methods55
III. Results56
IV. Discussion62
References66
Chapter 5. The Accumulation and Selective Detachment of Mitotic Cells71
I. Introduction71
II. Preparation of Mitotic Cells72
III. Colcemid and Mitotic Arrest73
IV. Cell Cycle AnaIysis76
V. Application to Other Cell Lines78
VI. Human Diploid Cells80
VII. Discussion81
References84
Chapter 6. Use of the Mitotic Selection Procedure for Cell Cycle Analysis: Emphasis on Radiation-Ind85
I. Introduction85
II. Materials and Methods87
III. Results92
IV. Discussion99
References101
Chapter 7. Evaluation of S Phase Synchronization by Analysis of DNA Replication in 5–Bromodeoxyuri103
I. Introduction103
II. Methods104
III. Results107
IV. Discussion110
References112
Chapter 8. Application of Precursors Adsorbed on Activated Charcoal for Labeling of Mammalian DNA in115
I. Introduction115
II. Preparation of Adsorbed Precursors116
III. Kinetics of Labeling of Mammalian DNA after a Single Injection of Adsorbed Thymidine-14C116
IV. Applications of the Technique118
V. Concluding Remarks121
References122
Chapter 9. Growth of Functional Glial Cells in a Serumless Medium123
I. Background Leading to Current Studies123
II. Composition, Preparation, and Storage of Synthetic Medium124
III. Adaptation and Care of Cells in Serumless Medium124
IV. Functional Characteristics of Adapted Cells128
V. Attempts to Grow Other Cells with Differentiated Characteristics in Serumless Medium136
VI. Significance of Studies and Future Directions136
References137
Chapter 10. Miniature Tissue Culture Technique with a Modified (Bottomless) Plastic Microplate139
I. Introduction140
II. Preparation of the Microplates140
III. Assembly of the Microplate for Use143
IV. Tissue Culture Procedures149
V. Use of the Plates150
VI. Removal of Slides153
VII. Staining of Slides154
VIII. Recycling the Plates155
References155
Chapter 11. Agar Plate Culture and Lederberg-Style Replica Plating of Mammalian Cells157
I. Introduction157
II. Agar Plate Culture158
III. Lederberg-Style Replica Plating of Mammalian Cells168
IV. Application of Agar Plate Culture and Replica Plating to Mammalian Cell Genetics171
V. Concluding Remarks176
References177
Chapter 12. Methods and Applications of Flow Systems for Analysis and Sorting of Mammalian Cells179
I. Introduction180
II. Operational Features of the Cell Analysis and Sorting System182
III. Light-Scattering Properties of Cells and Cell Models187
IV. Cell Preparative Procedures for Flow–System Analysis196
V. Methods for Staining Cells in Liquid Suspension202
VI. DNA Content Measurements of Diploid and Heteroploid Cells213
VII. CellCycle Analysis218
VIII. Analyses Employing Immunofluorescence Techniques230
IX. Determination of the Cell Surface Density Binding of Concanavalin A231
X. Applications of Cell-Sorting Techniques233
XI. Potential Applications for Fluorescent Enzyme Assay238
XII. Summary and Conclusions240
Appendix A: Cell Dispersal241
Appendix B: Acriflavin-Feulgen Procedure241
Appendix C: Ethidium Bromide (EB) and Propidium Iodide (PI) Protocols242
References242
Chapter 13. Purification of Surface Membranes from Rat Brain Cells247
I. The Neuronal Surface247
II. Isolation Methods249
III. Composition of Brain Plasma Membranes252
IV. Synthesis of Proteins and Glycoproteins of Surface Membranes254
References256
Chapter 14. The Plasma Membrane of KB Cells; lsolation and Properties259
I. Introduction260
II. Culture Conditions and Various Media261
III. Enzymic Assays261
IV. Analytical Methods262
V. Isolation of Plasma Membranes by the Latex Bead Method262
VI. Isolation of Plasma Membranes by the ZnCl2 Method263
VII. Isolation of Mitochondria and Microsomes from KB Cells264
VlII. Electron Microscopy265
IX. Kinetics of Bead Uptake266
X. Morphological Studies267
XI. Biochemical Studies272
XII. Kinetics of Transport of a-Aminoisobutyric Acid (AIB) in KB Cells After Ingestion of Latex Bead274
XIII. Purity of the Plasma Membranes274
XIV. Validity of the Latex Bead Method276
XV. Damage of the Plasma Membrane by Tris Buffer277
XVI. Applicability of the Latex Bead Method to Other Cells278
References278
Chapter 15. The Isolation of Nuclei from Paramecium aurelia281
I. Introduction281
II. Isolation of Macronuclei282
III. Isolation of Micronuclei298
IV. Perspectives308
References309
Chapter 16. Isolation of Micro- and Macronuclei of Tetrahymena pyriformiss311
I. Introduction311
II. Cell Culture313
Ill. Isolation of Nuclei315
IV. Yields and Purity of Isolated Nuclei320
V. Structural, Chemical, and Metabolic Studies of Isolated Nuclei325
References327
Chapter 17. Manipulations with Tetrahymena pyriformis on Solid Medium329
I. Introduction329
II. Growth and Cloning of Tetrahymena pyriformis on Solid Medium330
III. Summary and Conclusions347
References348
Chapter 18. The Isolation of Nuclei with Citric Acid and the Analysis of Proteins by Two-Dimensional349
I. Introduction349
II. Methods352
III. Morphology of Sucrose and Citric Acid Nuclei358
IV. Composition of Sucrose and Citric Acid Nuclei362
V. Two-Dimensional Electrophoretic Analysis of Nuclear Proteins from Citric Acid Nuclei362
VI. Isolation of Nucleoli373
VII. Summary374
References374
Chapter 19. Isolation and Manipulation of Salivary Gland Nuclei and Chromosomes377
I. Introduction377
II. Material378
III. Method380
IV. Dependence of Chromosome Structure on Ionic Environment387
References390
Index391

Book details

Vendor Elsevier S & T
SKU 9780125641098
ISBN-13 9780080859064
Author AUTHOR, UNKNOWN
Category Science
Subject Cell Biology

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METHODS IN CELL BIOLOGY,VOLUME 9

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