Note: Front Cover -- Methods in Cell Biology, Volume 61 -- Copyright Page -- Contents -- Contributors -- Preface -- Chapter 1. Isolation of Centrosomes from Drosophila Embryos -- I. Introduction -- II. Isolation of Centrosomes from Drosophila Embryos -- III. Assays for Microtubule Nucleation by Isolated Centrosomes -- IV. Conclusions -- References -- Chapter 2. Studying the Composition and Function of Centrosomes in Vertebrates -- I. Introduction -- II. Isolation of Centrosomes from Animal Cells -- III. Preparation of Immunological Probes from Isolated Centrosomes -- IV. Ultrastructural Analysis of Isolated Centrosomes -- V. Biochemical Composition of Centrosomes -- VI. Functional Assays of Isolated Centrosomes -- VII. Prospects -- References -- Chapter 3. Isolation of Centrosomes from Spisulu solidissima Oocytes -- I. Introduction -- II. Obtaining Organisms -- III. Isolation and Activation of Spisulu solidissima Oocytes -- IV. Preparation of Oocyte Lysates -- V. Preparation of Microtubule Protein -- VI. Isolation of Centrosomes from Activated Oocyte Lysates -- VII. lmmunofluorescence of Centrosomes and Asters -- VIII. Electron Microscopy of Asters and Centrosomes -- IX. Summary -- References -- Chapter 4. Methods for in Situ Localization of Proteins and DNA in the Centromere-Kinetochore Complex -- I. Introduction -- II. In Situ Localization of Proteins: Indirect Immunofluorescence -- III. In Situ Localization of Proteins: Immunogold EM -- IV. Fluorescent in Situ Hybridization Using DNA Satellite Probes -- V. Combination Staining: DNA/Protein -- VI. Specialized Techniques -- References -- Chapter 5. Three-Dimensional Transmission Electron Microscopy and Its Application to Mitosis Research -- I. Introduction -- II. Resolution and Choosing between Tomography and Serial Sections -- III. Electron Tomography -- IV. Serial Section Reconstruction. , V. Analysis and Display of 3D Reconstructions -- VI. Software Packages -- VII. Summary and Conclusions -- References -- Chapter 6. Enlightening Mitosis: Construction and Expression of Green Fluorescent Protein Fusion Proteins -- I. Introduction: Visualizing the Molecular Anatomy of the Spindle -- II. Fluorescence Properties of GFP -- III. Strategies for Constructing Fusion Proteins -- IV. Expression in Mammalian Cells -- References -- Chapter 7. Recombinant p50/Dynamitin as a Tool to Examine the Role of Dynactin in Intracellular Processes -- I. Introduction -- II. Production of Recombinant p50/Dynamitin -- III. Disruption of the Dynactin Complex by p50/Dynamitin in Xenopus Egg Extracts -- IV. Disruption of Spindle Poles Using p50/Dynamitin -- References -- Chapter 8. In Vitro Assays for Studying Saccharomyces cerevisiae Kinetochore Activity -- I. Introduction -- II. Microtubule-Binding Assays for S. cerevisiae Kinetochores -- Ill. Band Shift Assay for the Kinetochore Complex -- References -- Chapter 9. Fluorescent Speckle Microscopy of Spindle Microtubule Assembly and Motility in Living Cells -- I. lntroduction -- II. Principles of the Fluorescence Speckle Method for Microtubules -- III. Specimen Methods -- IV. Microscopy and Image Acquisition -- V. Image Processing and Analysis -- VI. Examples -- VII. Future Considerations -- References -- Chapter 10. Polarized Light Microscopy of Spindles -- I. Introduction -- II. Polarized Light Microscopy -- III. Analysis of Spindle Birefringence -- IV. Optimum Cell Types for Polarized Light Microscopy of Spindles -- References -- Chapter 11. Micromanipulation of Chromosomes and Spindles in Insect Spermatocytes -- I. Introduction -- II. Preparing for Micromanipulation -- Ill. Manipulating Cell Components -- References -- Chapter 12. Microinjection of Mitotic Cells -- I. lntroduction -- II. Choice of Cells. , III. Timing of Injection -- IV. Microinjection Procedure -- V. Conclusions -- References -- Chaspter 13. Obtaining Antibodies to Spindle Components -- I. Introduction -- II. Methods -- III. Discussion -- References -- Chapter 14. Using Antisense Technology to Study Mitosis -- I. Introduction -- II. Antisense Mechanism of Action -- III. Choice of Antisense Reagents -- IV. Assaying Target Protein Levels -- V. Antisense Reagents Used to Study Cell Division -- References -- Chapter 15. The Use and Action of Drugs in Analyzing Mitosis -- I. Introduction: Why Use Drugs? -- II. Brief Overview of Microtubule Assembly Dynamics -- III. Mechanisms of Action of Major Antimitotic Drugs: Binding to Tubulin and Microtubules and Effects on Microtubule Polymerization and Dynamics and on Mitosis -- IV. Determination of Intracellular Drug Levels -- V. How to Use Antimitotic Drugs: Practical Guidelines -- References -- Chapter 16. Correlative Light and Electron Microscopy of Mitotic Cells in Monolayer Cultures -- I. Introduction -- II. Light Microscopy -- III. Flat Embedding -- IV. Preparing the Cell for Sectioning -- V. Obtaining the Required Ultrastructural Information -- References -- Chapter 17. Identification and Characterization of Mitotic Mutations in Drosophila -- I. Introduction -- II. Maternal-Effect Mutations That Disrupt the Syncytial Mitotic Divisions -- III. Cytological Analysis of the Syncytial Mitoses -- IV. Zygotic Mutations That Disrupt Mitosis in Larval Tissues -- V. Cytological Analysis of Larval Brain and Imaginal Discs -- References -- Chapter 18. Methods for Isolating and Analyzing Mitotic Mutants in Aspergillus nidulans -- I. Introduction -- II. Characteristics of Aspergillus nidulans -- III. Strains and Media -- IV. Harvesting Conidia and Preparing Conidial Suspensions -- V. Mutagenesis. , VI. Methods for Isolating Mitotic Mutants in A . nidulans -- VII. Morphologcal Analysis of Mutants -- VIII. Genetic Analysis of Mutants -- IX. Molecular Genetic Methods for Working with A . nidulans -- References -- Chapter 19. Using Green Fluorescent Protein Fusion Proteins to Quantitate Microtubule and Spindle Dynamics in Budding Yeast -- I. Introduction -- II. Construction of Protein-GFP Fusion and Promoter Selection -- III. Quantifying Fluorescence in Cell Populations -- IV. The Imaging System -- V. Quantitative Solution to the Imaging Problem -- VI. lmage Acquisition and Processing -- VII. Applications and Examples: Expression of Dynein-GFP in Vivo -- References -- Chapter 20. The Use of Xenopus Egg Extracts to Study Mitotic Spindle Assembly and Function in Vitro -- I. Introduction -- II. Preparation of CSF Extracts for Spindle Assembly -- III. Spindle Assembly Reactions -- IV. Monitoring Spindle Assembly Reactions -- V. Manipulation of Extracts -- VI. Data Analysis and Interpretation -- VII. Anaphase in Vitro -- VIII. Conclusions -- References -- Chapter 21. Methods for Studying Cell Division in Higher Plants -- I. Introduction -- II. Cell Types of the Study of Cell Division -- III. Microinjection of Plant Cells -- IV. Conclusions -- References -- Chapter 22. Using Sea Urchin Gametes for the Study of Mitosis -- I. Introduction -- II. The Experimental System -- III. Maintenance of the Organisms -- IV. Obtaining Gametes -- V. Zygotes -- VI. Mounting Cells for Observation -- VII. Other Methods -- VIII. Annotated List of References -- References -- Index.