Note: Front Cover -- Methods in Cell Biology, Volume 59 -- Copyright Page -- Contents -- Contributors -- Preface -- PART I: Introduction -- Chapter 1. Overview of the Zebrafish System -- I. Introduction -- II. History of the Zebrafish System and Its Advantages and Disadvantages -- III. Cell and Developmental Biology, Organogenesis, and Human Disease -- IV. Genetics and Genomics -- V. Future Prospects -- VI. Conclusion -- References -- Chapter 2. Cell Cycles and Development in the Embryonic Zebrafish -- I. Introduction -- II. Terminology and the Staging Series -- III. The Zygote Period -- IV. The Cleavage Period -- V. The Blastula Period -- VI. The Gastrula Period -- VII. The Segmentation Period -- References -- PART II: Cell Culture and General Methods -- Chapter 3. Zebrafish Embryonal Cell Culture -- I. Introduction -- II. General Cell Culture Considerations -- III. Primary and Multipassage Zebrafish Embryo Cell Culture -- IV. Transfection -- V. Applications to Other Fish Models -- VI. Conclusion -- References -- Chapter 4. Primary Fibroblast Cell Culture -- I. Introduction -- II. Material and Methods -- III. Results and Discussion -- References -- Chapter 5. Production of Haploid and Diploid Androgenetic Zebrafish (Including Methodology for Delayed in Vitro Fertilization) -- I. Introduction -- II. Equipment and Materials -- III. Methods -- IV. Results and Discussion -- V. Conclusions and Perspectives -- References -- PART III. Gene Expression and Function in Development -- Chapter 6 . Analysis of Protein and Gene Expression -- I. In Situ Hybridlzation to RNA and Immunolocalization of Proteins -- II. Probe Synthesis -- III. Fixation -- IV. Hybridization to Whole-Mount Embryos -- V. Immunolocalization of Probes -- VI. Two-Color in Sit# Hybridization -- VII. Double-Fluorescent in Situ Hybridization. , VIII. Simultaneous Localization of Transcription and Translation Gene Products -- IX. Embedding and Sectioning Whole-Mount Embryos -- X. Solutions and Reagents -- References -- Chapter 7. Strategies to Perturb Zebrafish Development -- I. Introduction -- II. Administration of Reagents -- III. Reagents -- IV. Experiments to Modify the Activity of Specific Gene Products -- V. Perspectives: Inducible Systems -- References -- Chapter 8. Vectors and Techniques for Ectopic Gene Expression in Zebrafish -- I. Introduction -- II. Misexpression from DNA -- III. Misexpression from RNA -- References -- Chapter 9. Analysis of Zebrafish Development Using Explant Culture Assays -- I. Introduction -- II. Zebrafish Explants: General Considerations -- III. Materials Required -- IV. Guide to Explant Isolation and Culture -- V. Using Explants to Assay Induction -- VI. Illustrations of Specification and Induction Assays -- VII. Future Directions -- References -- PART IV: Early Embryonic Development -- Chapter 10. Embryonic Axis Formation in the Zebrafish -- I. The First Signs of Dorsal -- II. Establishment of Dorsal-Ventral Polarity in Blastula Stages -- III. Dorsal-Ventral Patterning -- IV. Development of Midline Mesodermal Tissues -- References -- Chapter 11. Confocal Microscopic Analysis of Morphogenetic Movements -- I. Introduction -- II. Confocal Imaging of Embryos -- III. General Principles of Vital Staining -- IV. Mounting Embryos for Imaging -- V. Imaging Procedures -- VI. Multilevel Time-Lapse Confocal Analysis -- VII. Distribution of Visual Information -- VIII. Confocal Imaging of Embryos Expressing Green Fluorescent Protein (GFP) -- IX. Summary -- References -- Chapter 12. Cytoskeletal Dynamics of the Zebrafish Embryo -- I. Introduction -- II. Cytoskeleton of the Unfertilized Egg -- III. Organization and Function of the Cytoskeleton in the Zygote. , IV. Cleavage and Blastula Period -- V. Yolk Cell Microtubules during Epiboly -- VI. Tubulin Dynamics in Neuronal Axons of Living Zebrafish Embryos -- VII. Methods -- References -- Chapter 13. Kinesin-like Microtubule Motors in Early Development -- I. Introduction -- II. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Cloning Strategy for Zebrafish Kinesin-like Protein Genes (KLPs) -- III. Production of Anti-zIUP Antibodies -- IV. Expression of GFP-Tagged zKLPs in Cultured Cells -- V. General Considerations -- References -- PART V: Organogenesis -- Chapter 14. Techniques in Neural Development -- I. Introduction -- II. Organizers of Anterior-Posterior Pattern in the Neural Plate -- III. Zebrafish Mutations That Disrupt A-P Pattern in the Neural Tube -- IV. Techniques for the Study of Neural Patterning Mutants -- Appenhx: Codetection of Lineage Marker and Gene Expression in Whole-Mount Embryos -- References -- Chapter 15. Development of the Retina -- I. Introduction -- II. Development of the Zebrafish Retina -- IV. Genetic Analysis of the Zebrafish Retina -- V. Summary -- References -- Chapter 16. Growth Control in the Ontogenetic and Regenerating Zebrafish Fin -- I. Introduction -- II. Mutations Affect the Growth, Development, and Regeneration of Fins -- III. Developmental Checkpoints in Fin Growth -- IV. Cellular Basis of Growth in the Fin -- V. Conclusion -- References -- Chapter 17. Vascular and Blood Gene Expression -- I. Introduction -- II. Embryonic Vascularization -- III. Embryonic Hematopoiesis -- IV. Vascular and Hematopoietic Gene Expression during Development -- V. Mutations Affecting Vascularization and Hematopoiesis -- VI. Conclusion and Future Directions -- References -- Chapter 18. Analysis of Hemostasis in the Zebrafish -- I. Introduction -- II. Methods -- III. Future Assay Development and Applications -- References. , Chapter 19. Cell Lineage Tracing in Heart Development -- I. Introduction -- II. Methodology -- III. Analysis -- IV. Summary -- References -- Chapter 20. Neurogenesis in Zebrafish Embryos -- I. The System -- II. Early Zebrafish Embryos Have a Simple Nervous System -- III. Origins and Morphogenesis of the Nervous System -- IV. Antagonists of Bone Morphogenetic Protein (BMP) Activity Lead to a Neural Fate in the Ectoderm -- V. Fibroblast Growth Factor-8 (FGF-8) Defines Dorsoventral Positional Information in the Ectoderm -- VI. Interactions with the Germ-Ring Provide Posteriorizing Signals -- VII. Ablation of Cells in the Anterior Ectoderm Identifies a Novel Organizer -- VIII. Patterning of the Neural Plate by Midline Signals: The Role of Factors in the Hedgehog, EGF, and TGF-0 Families -- IX. BMPs Induce Dorsal Neurons -- X. Controlling the Number of Neurons: Proneuronal Domains and Proneural and Neurogenic Genes -- XI. Position with Respect to Somite Boundaries Determines Motorneuron Subtype -- XII. Patterning Neurogenesis in the Epiphysial Neurons -- XIII. valentino (val) Suggests That Rhombomere Organization Is Preceded by the Fonnation of "Protosegments" -- XIV. Perspectives -- References -- Index -- Color Plate Section.

Note: Front Cover -- Methods in Cell Biology, Volume 60 -- Copyright Page -- Contents -- Contributors -- Preface -- Chapter 1. A Gynogenesis-Based Screen for Maternal-Effect Genes in the Zebrafish, Danio rerio -- I. Introduction -- II. Optimization of the Gynogenetic Method -- III. A Pilot Screen for Maternal-Effect Mutations -- IV. Recovery of Mutations -- V. Solutions, Materials, and Protocols for Gynogenetic Methods -- VI. Conclusions -- References -- Chapter 2. Developmental Mutant Screens in the Zebrafish -- I. Introduction -- II. Mutagenesis -- III. Breeding Scheme -- IV. Screening Procedure -- V. Allele Screens -- VI. Dominant Screens -- VII. Managing the Outcome of Screens -- VIII. Potential of Future Screens -- Appendix-Recipes and Media -- References -- Chapter 3. Haploid Screens and Gamma-Ray Mutagenesis -- I. Introduction -- II. Production of Haploid Embryos -- III. Development of Haploid Embryos -- IV. Genetic Background -- V. The Haploid Screen -- VI. Limitations of Haploids -- VII. Mosaicism in F1 Screens -- VIII. Haploid Screens and Gamma-Ray Mutagenesis -- IX. Nature of Gamma-Ray-Induced Mutations -- X. Conclusion -- References -- Chapter 4. Early Pressure Screens -- I. Introduction -- II. Rationale for Use of Early Pressure to Produce Diploid Embryos for Genetic Screens -- III. Rationale for Use of Screens Focused on Specific Phenotypes -- IV. Design of Our Screen and Mutant Phenotypes Isolated -- V. Closing Remarks -- References -- Chapter 5. Retrovirus-Mediated Insertional Mutagenesis in Zebrafish -- I. Introduction -- II. Insertional Mutagenesis: A Pilot Screen -- III. Considerations for a Large-Scale Screen -- References -- Chapter 6. Genetic Applications of Transposons and Other Repetitive Elements in Zebrafish -- I. Introduction -- II. Classification of Repetitive Elements -- III. Genetic Applications of Repetitive Elements. , IV. Summary and Perspectives -- References -- Chapter 7. Transgenesis -- I. Introduction -- II. Materials -- III. Methods -- IV. Discussion -- References -- Chapter 8. The Zebrafish Genome -- I. Introduction -- II. Mapping the Zebrafish Genome -- III. The Zebrafish Gene Map -- IV. Two Genes in Zebrafish for One in Mammals -- V. Gene Nomenclature in Zebrafish -- VI. Conclusions -- References -- Chapter 9. Using Random Amplified Polymorphic DNAs in Zebrafish Genomic Analysis -- I. Introduction -- II. What Are RAPDs? -- III. Advantages and Disadvantages of RAPDs as Polymorphic Markers -- IV. Methods for Developing RAPD Markers -- V. Using RAPDs to Identify Markers Closely Linked to a Mutation -- VI. Conversion of RAPD Markers to Sequence Tagged Sites -- VII. Conclusions -- References -- Chapter 10. Simple Sequence-Length Polymorphism Analysis -- I. Analysis in DNA Sequencing Gels -- II. Alternative Analysis of SSLP-Containing PCR Product in High-Resolution Agarose Gels -- III. Troubleshooting and Other Considerations -- References -- Chapter 11. Gene Mapping in Zebrafish Using Single-Strand Conformation Polymorphism Analysis -- I. Introduction -- II. SSCP Methodology -- III. Gene Mapping -- IV. Methods -- References -- Chapter 12. Mapping Zebrafish Mutations by AFLP -- I. Introduction -- II. Extraction of Zebrafish Genomic DNA -- III. AFLP Methods -- IV. General Considerations -- References -- Chapter 13. Zebrafish Expressed Sequence Tags and Their Applications -- I. Introduction -- II. Procedure of cDNA Clone Tagging -- III. Management of Tagged cDNA Clones -- IV. Applications of Tagged cDNA Clones -- References -- Chapter 14. Zebrafish YAC, BAC, and PAC Genomic Libraries -- I. Introduction -- II. Cloning Systems -- III. Available Zebrafish Genomic Libraries and the Need for Multiple Libraries -- IV. Screening Methods. , V. Positional Cloning Using YAC, BAC, and PAC Libraries -- VI. Further Applications of YAC, BAC, and PAC Resources to Zebrafish Genetics -- VII. Summary -- Internet Web Sites Relevant to YAC, BAC, and PAC Cloning -- References -- Chapter 15. Positional Cloning of Mutated Zebrafish Genes -- I. Introduction -- II. Initiating a Positional Cloning Project -- III. Isolating Genomic Clones and Defining the Critical Region -- IV. Gene Discovery -- References -- Chapter 16. Construction and Characterization of Zebrafish Whole Genome Radiation Hybrids -- I. Introduction -- II. Basic Protocol: Construction of Radiation Hybrids -- III. Characterization of Radiation Hybrids by PCR -- IV. Characterization of Radiation Hybrids Using FISH Analysis -- V. Commentary -- VI. Critical Parameters and Troubleshooting -- References -- Chapter 17. Zebrafish/Mouse Somatic Cell Hybrids for the Characterization of the Zebrafish Genome -- I. Introduction -- II. Production of Zebrafish/Mouse Somatic Cell Hybrids -- III. Analysis of Zebrafish Chromosomes in Hybrid Cells by Fluorescence in Situ Hybridization -- IV. Identification of Chromosomes in Hybrids Using PCR -- V. Inter-DANA/Mermaid PCR on Hybrids -- VI. Stability of Zebrafish/Mouse Hybrids -- VII. Perspectives -- References -- Chapter 18. Banded Chromosomes and the Zebrafish Karyotype -- I. Introduction -- II. Methods -- III. Results and Discussion -- References -- Chapter 19. Zebrafish Informatics and the ZFIN Database -- I. Introduction -- II. Database Design Process -- III. The Data Model and the Contents of the Database -- IV. Using the ZFIN Database -- References -- Appendix 1. Genetic Backgrounds and Some Standard Stocks and Strains Used in Zebrafish Developmental Biology and Genetics -- Summary of Genetic Backgrounds -- References -- Appendix 2. Centromeric Markers in the Zebrafish -- References. , Appendix 3. Collection, Storage, and Use of Zebrafish Sperm -- I. Introduction -- II. Materials -- III. Collection of Zebrafish Sperm -- IV. Cryopreservation of Zebrafish Sperm -- V. In Vitro Fertilization -- VI. General Considerations -- References -- Appendix 4. Zebrafish Web Site Listings -- Zebrafish Web Servers -- Lab URLs -- Index -- Color Plate Sections.

Note: Front Cover -- Methods in Cell Biology: Cumulative Subject Index -- Copyright Page -- Contents -- Cumulative Subject Index -- Contributor Index.

Note: Front Cover -- Methods in Cell Biology, Volume 47 -- Copyright Page -- Contents -- Contributors -- Preface -- Introduction -- Part I: Isolation of Cilia and Flagella -- Chapter 1. Flagellar Amputation and Regeneration in Chlamydomonas -- I. Introduction -- II. Procedures for Flagellar Amputation -- III. Assay of Flagellar Regeneration -- References -- Chapter 2. Large-Scale Isolation of Chlamydomonas Flagella -- I. Introduction -- II. Methods -- References -- Chapter 3. Isolation of Cilia from Tetrahymena thermophila -- I. Introduction -- II. Tetrahymena Cell Culture -- III. Isolation of Tetrahymena Cilia -- References -- Chapter 4. Preparation of Cilia and Subciliary Fractions from Paramecium -- I. Introduction -- II. Solutions -- III. Methods of Deciliation -- IV. Preparation of Subciliary Fractions -- References -- Chapter 5. Isolation of Euglena Flagella -- I. Introduction -- II. Cell Culture -- III. Isolation of Flagella -- References -- Chapter 6. Isolation of Ram Sperm Flagella -- I. Introduction -- II. Preparation of Ram Sperm Flagella -- III. Demembranation of Ram Sperm Flagella -- References -- Chapter 7. Isolation of Molluscan Gill Cilia, Sperm Flagella, and Axonemes -- I. Introduction -- II. Biological Material -- III. Methods -- References -- Chapter 8. Isolation of Flagella and Their Membranes from Sea Urchin Spermatozoa -- I. Introduction -- II. Experimental Procedures -- References -- Chapter 9. Isolation of Fish Sperm Flagella -- I. Introduction -- II. Media -- III. Collection of Sperm -- IV. Isolation of Axonemes -- V. Isolation of Flagella -- VI. Demembranation and ATP Reactivation of Isolated Carp Sperm Flagella -- VII. Conclusion -- References -- Part II: Culture and Isolation of Ciliated Epithelial Cells and Cilia -- Chapter 10. Ciliogenesis of Rat Tracheal Epithelial Cells in Vitro -- I. Introduction. , II. Isolation and Culture of Cells -- III. Comments and Conclusions -- References -- Chapter 11. Preparation of Explant and Organ Cultures and Single Cells from Airway Epithelium -- I. Introduction -- II. Methods -- References -- Chapter 12. Induction of Ciliogenesis in Oviduct Epithelium -- I. Introduction -- II. Methods -- III. Troubleshooting -- References -- Chapter 13. Preparation of Teleost Rod Inner and Outer Segments -- I. Introduction -- II. Method -- References -- Chapter 14. Isolation of Respiratory Cilia -- I. Introduction -- II. Materials -- III. Procedure -- References -- Part III: General Methods -- Chapter 15. Electrophoretic Separation of Dynein Heavy Chains -- I. Introduction -- II. Chemicals, Solutions, and Apparatus -- III. Procedure -- IV. Pitfalls -- References -- Chapter 16. Two-Dimensional Separation of Axonemal Proteins -- I. Introduction -- II. Chemicals, Solutions, and Apparatus -- III. Procedure -- IV. Advantages -- V. Pitfalls -- References -- Chapter 17. Analysis of Flagellar Calcium-Binding Proteins -- I. Background and Rationale -- II. Materials -- III. Diagonal Gel Procedures -- References -- Chapter 18. Phosphorylation of Chlamydomonas Flagellar Proteins -- I. Introduction -- II. In Vivo Labeling of Chlamydomonas Flagellar Proteins with [32P]-Orthophosphoric Acid -- References -- Chapter 19. In Situ Glycosylation of Flagellar Lipids -- I. Introduction -- II. Protocol for Demonstrating Glucosyltransferase Activity -- III. Glucosyltransferase(s): Enrichment and Identification of Polyacrylamide Gels -- References -- Chapter 20. Detection of Flagellar Protein Kinases on Polyvinylidene Fluoride Membranes Following Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis -- I. Introduction -- II. Methods -- References -- Chapter 21. Measurement of ATPase Activity Using [γ-32P]ATP -- I. Introduction -- II. Methods. , References -- Chapter 22. Nonradioactive Method for ATPase Assays -- I. Introduction -- II. Method -- References -- Part IV: Structure -- Chapter 23. Immunoelectron Microscopy -- I. Introduction -- II. Primary Antibodies -- III. Specimen Preparation -- IV. Immunolabeling Technique -- References -- Chapter 24. Immunofluorescence Microscopy of Cilia and Flagella -- I. Introduction -- II. General Considerations -- III. Reagents and Solutions -- IV. Immunofluorescence of Ciliated and Flagellated Cells -- V. Troubleshooting and Variations -- References -- Chapter 25. Scanning Transmission Electron Microscopic Analysis of the Isolated Dynein ATPase -- I. Introduction -- II. Method -- References -- Chapter 26. High-Resolution Negative Staining of the Isolated Dynein ATPase -- I. Introduction -- II. Method -- References -- Chapter 27. Computer-Assisted Analysis of Flagellar Structure -- I. Introduction -- II. Specimen Preparation and Electron Microscopy -- III. Image Analysis -- IV. Quantitative Analysis -- References -- Chapter 28. Preparation of Cilia and Flagella for Thin-Section Transmission Electron Microscope Analysis -- I. Introduction -- II. Sources of Materials -- III. Procedures -- References -- Chapter 29. Fixation of Mammalian Spermatozoa for Electron Microscopy -- I. Introduction -- II. Choice of Fixative -- III. Fixation of Sperm in Situ -- IV. Fixation of Sperm in Suspension -- V. Conclusion: A Few Hints -- Reference -- Part V: Motors and Motion Analysis -- Chapter 30. Reactivation of Chlamydomonas Cell Models -- I. Introduction -- II. Methods -- III. Comments -- References -- Chapter 31. Reactivation of Extracted Paramecium Models -- I. Introduction -- II. Methods -- References -- Chapter 32. Isolation and Reactivation of Newt Lung Respiratory Cilia -- I. Introduction -- II. Methods -- References. , Chapter 33. Reactivation of Motility of Demembranated Sea Urchin Sperm Flagella -- I. Introduction -- II. General Considerations -- III. Methods -- IV. Comments -- V. Extensions to Other Species -- VI. Algorithm for Calculating Recipes for Solutions -- VII. Health and Safety Considerations -- References -- Chapter 34. High-Speed Video Microscopy of Flagella and Cilia -- I. Introduction -- II. Video Cameras -- III. Other Equipment -- IV. Procedure for Recording -- References -- Chapter 35. Micromanipulation of Sperm and Other Ciliated or Flagellated Single Cells -- I. Introduction -- II. Methods -- References -- Chapter 36. Preparation and Reactivation of Demembranated, Cytosol-Free Ram Spermatozoa -- I. Introduction -- II. Methods -- References -- Chapter 37. Microscopic Assays of Flagellar Dynein Activity -- I. Introduction -- II. Methods -- References -- Chapter 38. High-Resolution Imaging of Flagella -- I. Introduction -- II. Methods -- III. Conclusion -- References -- Chapter 39. Flagellar Surface Motility: Gliding and Microsphere Movements -- I. Introduction -- II. Description of Gliding Motility -- III. Procedures -- IV. Nongliding Mutants -- V. Perspectives -- References -- Chapter 40. Assay of Chlamydomonas Phototaxis -- I. Introduction -- II. Culture and Handling of Chlamydomonas Cells -- III. Phototaxis Room Setup -- IV. Equipment for Actinic Illumination of Cells during Assay -- V. Phototaxis Chamber -- VI. Microscopy -- VIl. Video Equipment -- VIII. Analysis and Presentation -- References -- Chapter 41. Quantification of Ciliary Beat Frequency and Metachrony by High-Speed Digital Video -- I. Introduction -- II. Optimization of the Light Signal Waveform Produced by Beating Cilia -- III. Quantification of Ciliary Metachronism -- IV. Video Techniques for Measuring Ciliary Activity -- V. A High-Speed Digital Video Photodetection Technique. , VI. Summary -- References -- Part VI: Cytoskeletal Preparations -- Chapter 42. Isolation and Fractionation of the Tetrahymena Cytoskeleton and Oral Apparatus -- I. Introduction -- II. Methods -- References -- Chapter 43. Release of the Cytoskeleton and Flagellar Apparatus from Chlamydomonas -- I. Introduction -- II. Strains and Reagents -- III. Solutions -- IV. Procedures -- V. Additional Treatments -- References -- Chapter 44. Purification of SF-Assemblin -- I. Introduction -- II. Methods -- III. Comments -- References -- Chapter 45. Purification of Basal Bodies and Basal Body Complexes from Chlamydomonas reinhardtii -- I. Introduction -- II. Methods -- III. Comparison of Protocols -- IV. Components of Basal Body Complexes -- V. Learning about Function -- References -- Chapter 46. Preparation and Reactivation of Spermatozopsis Cytoskeletons -- I. Introduction -- II. Methods -- References -- Chapter 47. Centrin-Based Contractile Fibers: Chromatographic Purification of Centrin -- I. Introduction -- II. Methods -- III. Perspectives -- References -- Part VII: Flagellar Fractionation and Membrane Characterization -- Chapter 48. Isolation of Flagellar Paraxonemal Rod Proteins -- I. Introduction -- II. Methods -- References -- Chapter 49. Preparation of Ciliary and Flagellar Remnants -- I. Introduction -- II. Remnant Fractionation -- References -- Chapter 50. In Vitro Polymerization of Tubulin from Echinoderm Sperm Flagellar Microtubules -- I. Introduction -- II. Methods -- III. Discussion -- References -- Chapter 51. Methods for the Isolation of Tektins and Sarkosyl-Insoluble Protofilament Ribbons -- I. Introduction -- II. Methods -- III. Discussion -- References -- Chapter 52. Isolation of Radial Spoke Heads from Chlamydomonas Axonemes -- I. Introduction -- II. Method -- References -- Chapter 53. Isolation of the Dense Fibers of Mammalian Sperm Flagella. , I. Introduction.

Note: Front Cover -- Methods in Cell Biology, Volume 49 -- Copyright Page -- Contents -- Contributers -- Preface -- Part I: Techniques for Examination of Cells within Tissues -- Chapter 1. Advances in High-pressure and Plunge-Freeze Fixation -- I. Introduction -- II. Plunge-Freeze Fixation -- III. High-pressure Freeze Fixation -- IV. Results and Discussion -- V. Conclusions and Perspectives -- References -- Chapter 2. Ion Localization and X-Ray Microanalysis -- I. Introduction -- II. Plant Materials -- III. The Choice of Methods -- IV. Cryo-SEM and X-Ray Microanalysis -- V. Freeze Substitution and TEM -- VI. Other Preparative Techniques -- VII. Other Machinery Used for Ion Localization -- VIII. Conclusions -- References -- Chapter 3. Freeze-Fracture Deep-Etch Methods -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedure -- V. Conclusions -- References -- Chapter 4. Advances in Immunoelectron Microscopy -- I. Introduction -- II. Tissue Preparation -- III. On-Grid Section Labeling -- IV. Protocols -- V. Conclusions and Perspectives -- References -- Chapter 5. Freeze-Substitution -- I. Introduction -- II. Freeze-Substitution -- III. Conclusion and Perspectives -- References -- Chapter 6. Cell Optical Displacement Assay (CODA) -Measurements of Cytoskeletal Tension in Living Plant Cells with a Laser Optical Trap -- I. Introduction -- II. Optical Trapping-A Tool for Cellular Measurements and Manipulations -- III. Cell Optical Displacement Assay (CODA)-A Quantitative Tool for Tension Analysis in the Cytoskeleton of Living Plant Cells -- IV. CODA and the Plant Cell-Conclusions -- References -- Chapter 7. Methods in Plant Immunolight Microscopy -- I. Introduction -- II. Preparing Isolated Cells for Immunostaining -- III. Immunostaining -- IV. Mounting -- V. Image Recording. , VI. Sectioning Intact Tissues for Immunostaining -- VII. Perspectives -- References -- Chapter 8. Confocal Epipolarization Microscopy of Gold Probes in Plant Cells and Protoplasts -- I. Introduction -- II. Configuring a Confocal Laser Scanning Microscope for Epipolarization -- III. Imaging Colloidal Gold in Protoplasts -- IV. Imaging Biolistically Delivered Gold Beads -- V. Conclusions -- References -- Chapter 9. Monoclonal Antibodies to Cell-Specific Cell Surface Carbohydrates in Plant Cell Biology and Development -- I. Applications -- II. Methods -- III. Perspectives -- References -- Chapter 10. Epitope Tagging for the Detection of Fusion Protein Expression in Transgenic Plants -- I. Introduction -- II. Methods -- III. Results and Discussion -- IV. Conclusions and Perspectives -- References -- Chapter 11. In Situ Enzyme Histochemistry on Plastic-Embedded Plant Material -- I. Introduction -- II. Methods -- III. Results and Discussion -- IV. Conclusion -- References -- Chapter 12. In Situ Hybridization -- I. Introduction -- II. Which Protocol to Use? -- III. Labels, Tags, and Detection Systems -- IV. Probe Types -- V. Hybridization Conditions -- VI. Controls -- VII. New Techniques -- References -- Chapter 13. Localization of RNA by High Resolution in Situ Hybridization -- I. Introduction -- II. Principles of in Situ Hybridization Technique at the Electron Microscopy Level -- III. Materials -- IV. Methods -- V. Results and Discussion -- VI. Perspectives -- References -- Chapter 14. Recombinant Aequorin Methods for Intracellular Calcium Measurement in Plants -- I. Introduction -- II. Expression of Recombinant Aequorin in Plants -- III. Reconstitution of Aequorin -- IV. Measurement of Light -- V. Perspectives -- References -- Chapter 15. Confocal Microscopy of the Shoot Apex -- I. Introduction -- II. Materials -- III. Methods. , IV. Results and Discussion -- V. Conclusions and Perspectives -- References -- Chapter 16. Measurements of Wall Stress Relaxation in Growing Plant Cells -- I. Introduction -- II. Theory Underlying the Method -- III. Materials -- IV. Methods -- V. Critical Aspects of the Procedure -- VI. Results and Interpretation -- VII. Conclusion and Perspectives -- References -- Chapter 17. High-Resolution NMR Methods for Study of Higher Plants -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedures -- V. Results and Discussion -- VI. Conclusions and Perspectives -- References -- Chapter 18. Electrophysiology -- I. Introduction -- II. Materials and Methods -- III. Critical Aspects of the Procedure -- IV. Results and Discussion -- V. Conclusion and Perspectives -- References -- Chapter 19. Ion-Selective Microelectrodes for Measurement of Intracellular Ion Concentrations -- I. Introduction -- II. Materials -- III. Methods -- IV. Results -- V. Conclusions and Perspectives -- References -- Chapter 20. Patch-Clamping Plant Cells Patch-Clamping Plant Cells -- I. Introduction -- II. Materials and Methods -- III. Results and Discussion -- References -- Part II: Techniques for Manipulation and Analysis of Different Cell Types -- Chapter 21. Methods for Mesophyll and Bundle Sheath Cell Separation -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedure -- V. Results and Discussion -- VI. Conclusion and Perspectives -- References -- Chapter 22. Synchronization of Cell Cultures of Higher Plants -- I. Introduction -- II. Application -- III. Plant Materials -- IV. Procedures -- V. Critical Aspects for Synchronization -- VI. Results -- VII. Discussion -- References -- Chapter 23. Genetic Tagging of Cells and Cell Layers for Studies of Plant Development -- I. Introduction -- II. Chimeras. , III. Spontaneous Sectors -- IV. Sectors Induced at Specific Stages of Development -- V. Sector Boundary Analysis -- VI. Pattern Determination -- VII. Perspectives -- References -- Chapter 24. Chemically Induced Mitotic Synchrony in Root Apical Meristems -- I. Introduction -- II. Materials and Recipes -- III. Aspects of the Procedure -- IV. Methods -- V. Perspectives -- References -- Chapter 25. Manipulation of Pollen Grains for Gametophytic and Sporophytic Types of Growth -- I. Introduction -- II. Materials and Methods -- III. Critical Aspects of the Procedures -- IV. Conclusions and Perspectives -- References -- Chapter 26. Root Border Cells as Tools in Plant Cell Studies -- I. Introduction -- II. Materials -- III. Methods -- IV. Special Considerations -- V. Applications of Border Cells in Teaching and Research -- VI. Summary and Perspectives -- References -- Part III: Signal Transduction and Information Transfer -- Chapter 27. In Vivo Footprinting of Protein-DNA Interactions -- I. Introduction -- II. Materials -- III. Methods -- IV. Typical Results and Discussion -- V. Conclusions and Perspectives -- References -- Chapter 28. The Interaction Trap: In Vivo Analysis of Protein-Protein Associations -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedure -- V. Conclusions and Perspectives -- References -- Chapter 29. Cloning Plant Genes by Complementation of Yeast Mutants -- I. Introduction -- II. Materials and Methods -- III. Critical Aspects of the Procedures -- IV. Results and Discussion -- V. Conclusion and Perspectives -- References -- Chapter 30. Differential mRNA Display -- I. Introduction -- II. Methods -- III. Critical Aspects of the Technique -- IV. Potential Modifications of the Procedure -- V. Results and Discussion -- VI. Conclusions and Perspectives -- References. , Chapter 31. Molecular Methods for Isolation of Signal Transduction Pathway Mutants -- I. Introduction -- II. Method I: Transformation of Arabidopsis thaliana -- III. Method II: Screen/Selection for Signal Transduction Pathway Mutants -- IV. Genetic Characterization -- V. Results and Discussion -- VI. Conclusions -- References -- Chapter 32. Induction of Signal Transduction Pathways through Promoter Activation -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedure -- V. Discussion -- References -- Chapter 33. In Vitro Analysis of G-Protein Functions -- I. Introduction -- II. Materials -- III. Methods for Detecting G Proteins -- IV. Analysis of G-Protein Involvement in Cellular Processes -- V. Conclusion -- References -- Chapter 34. Production of Recombinant Plant Calmodulin and Its Use to Detect Calmodulin-Binding Proteins -- I. Introduction -- II. Materials -- III. Methods -- IV. Critical Aspects of the Procedures -- V. Results -- References -- Chapter 35. Analysis of the Light Signaling Pathway in Stomatal Guard Cells -- I. Introduction -- II. Materials and Methods -- III. Results and Discussion -- IV. Conclusions and Perspectives -- References -- Chapter 36. Immunocytochemical Localization of Receptor Protein Kinases in Plants -- I. Introduction -- II. Immunolocalization in Paraffin-Embedded Tissue by Light Microscopy -- III. Immunolocalization in Plastic-Embedded Tissue by Light Microscopy -- IV. Immunolocalization by Electron Microscopy -- V. Formulations -- VI. Conclusions -- References -- Chapter 37. Expression and Assay of Autophosphorylation of Recombinant Protein Kinases -- I. Introducton -- II. Materials -- III. Selecting the Region of a Protein Kinase Molecule to Express -- IV. Choosing the Best Expression Vector -- V. Choice of E. coli Strains for Heterologous Protein Expression. , VI. Isolation of Fusion Proteins.

Note: Front Cover -- Methods in Cell Biology, Volume 41 -- Copyright Page -- Contents -- Contributors -- Preface to the Second Edition -- Preface to the First Edition -- Chapter 1. Dissociation of Intact Cells from Tumors and Normal Tissues -- I. Introduction -- II. Application -- III. Methodology: Mechanical Cell Dispersion -- IV. Results -- V. Conclusion -- References -- Chapter 2. Assays of Cell Viability: Discrimination of Cells Dying by Apoptosis -- I. Introduction -- II. Changes in Light Scatter during Cell Death -- III. Cell Sensitivity to Trypsin and DNase -- IV. Fluorescein Diacetate (FDA) Hydrolysis and PI Exclusion -- V. Rh123 Uptake and PI Exclusion -- VI. PI Exclusion Followed by Counterstaining with Hoechst 33342 -- VII. Hoechst 33342 Active Uptake and PI Exclusion -- VIII. Controlled Extraction of Low MW DNA from Apoptotic Cells -- IX. Sensitivity of DNA in Situ to Denaturation -- X. Detection of DNA Strand Breaks in Apoptotic Cells -- XI. A Selective Procedure for DNA Extraction from Apoptotic Cells Applicable to Gel Electrophoresis and Flow Cytometry -- XII. Comparison of the Methods: Confirmation of the Apoptotic Mode of Cell Death -- References -- Chapter 3. Cell Preparation for the Identification of Leukocytes -- I. Introduction -- II. Antibodies -- III. Titering Antibodies -- IV. Third Color Reagents -- V. Cell Preparation -- VI. Labeling Procedures -- VII. Lysing Erythrocytes -- VIII. Detecting Dead Cells -- IX. Fixation -- X. Solutions and Reagents -- References -- Chapter 4. Multiparameter Analysis of Leukocytes by Flow Cytometry -- I. Introduction -- II. Correlated List-Mode Data -- III. Single-Label Analysis -- IV. Optical Filtration -- V. Compensation -- VI. Two-Color Fluorescence -- VII. Three-Color Immunophenotyping -- VIII. Fixing Cells -- IX. Detecting Dead Cells -- X. Summary -- References. , Chapter 5. Immunophenotyping Using Fixed Cells -- I. Introduction -- II. Application -- III. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Results and Discussion -- References -- Chapter 6. Simultaneous DNA Content and Cell-Surface Immunofluorescence Analysis -- I. Introduction -- II. Materials and Methods -- III. Instruments -- IV. Critical Aspects of the Method -- V. Results -- VI. Applications and Future Directions -- References -- Chapter 7. Flow Cytometry Crossmatching for Solid Organ Transplantation -- I. Introduction -- II. Applications -- Ill. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Standards -- VII. Instruments -- VIII. Results and Discussion -- References -- Chapter 8. Cell Membrane Potential Analysis -- I. Introduction -- II. Materials and Methods -- III. Improving Cytometry of Membrane Potentials -- References -- Chapter 9. Measurement of Intracellular pH -- I. Introduction -- II. Application -- Ill. Materials -- IV. Cell Preparation and Staining: BCECF or SNARF -- V. lnstruments -- VI. Critical Aspects -- VII. Results and Discussion -- References -- Chapter 10. Intracellular Ionized Calcium -- I. Introduction -- II. Flow Cytometric Assay With Indo-1 -- III. Use of Flow Cytometry and Fluo-3 to Measure [Ca2+]i -- IV. Use of Flow Cytometry and Fluo-3/Fura-Red for Ratiometric Analysis of [Ca2+]i -- V. Cell Conjugate Assays Combined with Calcium Analysis and Flow Cytometry -- VI. Pitfalls and Critical Aspects -- VII. Limitations -- VIII. Results -- References -- Chapter 11. Cellular Protein Content Measurements -- I. lntroduction -- II. Applications -- III. Materials -- IV. Cell Preparation and Staining -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results and Discussion -- References. , Chapter 12. Lysosomal Proton Pump Activity: Supravital Cell Staining with Acridine Orange Differentiates Leukocyte Subpopulations -- I. Introduction -- II. Applications -- III. Materials -- IV. Staining Procedures -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- References -- Chapter 13. Staining of DNA in Live and Fixed Cells -- I. Introduction -- II. Applications with DNA-Specific Fluorochromes -- III. Materials -- IV. Cell Preparation and Fixation -- V. Cell Staining -- VI. Critical Aspects -- VII. Controls and Standards -- VIII. Instruments -- IX. Results and Discussion -- References -- Chapter 14. High-Resolution Analysis of Nuclear DNA Employing the Fluorochrome DAPI -- I. Introduction -- II. Applications -- III. Materials -- IV. Cell Separation and Fixation Method -- V. Staining Procedure -- VI. Instrument Setup -- VII. Standards -- VIII. Results and Discussion -- References -- Chapter 15. Detergent and Proteolytic Enzyme-Based Techniques for Nuclear Isolation and DNA Content Analysis -- I. Introduction -- II. Basic Principles of the Methods -- III. Applications -- IV. Materials -- V. Methods -- VI. Results -- References -- Chapter 16. DNA Analysis from Paraffin-Embedded Blocks -- I. Introduction -- II. Application -- III. Methods -- IV. Critical Aspects of Technique -- V. Alternative Methods for Sample Preparation -- References -- Chapter 17. Controls, Standards, and Histogram Interpretation in DNA Flow Cytometry -- I. Introduction -- II. Application -- III. Instrument Quality Control and Standardization -- IV. DNA Histogram Interpretation -- V. Methods -- VI. Summary of Criteria for Interpretation of DNA Histograms -- References -- Chapter 18. DNA Content Histogram and Cell-Cycle Analysis -- I. Introduction -- II. DNA Content Histogram Basic Principles. , III. Cell-Cycle Analysis of DNA Content Histograms -- IV. Critical Aspects of DNA Content and Cell-Cycle Analysis -- V. Interpretation of DNA Content and Cell-Cycle Histograms -- VI. Conclusion -- References -- Chapter 19. Imniunochemical Quantitation of Bromodeoxyuridine: Application to Cell-Cycle Kinetics -- I. Introduction -- II. Applications -- III. Materials -- IV. Procedures -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- IX. Summary -- References -- Chapter 20. Application and Detection of IdUrd and CldUrd as Two Independent Cell-Cycle Markers -- I. Introduction -- II. Application -- III. Materials -- IV. Methods -- V. Critical Aspects of the Procedure -- VI. Controls and Standards -- VII. Results -- References -- Chapter 21. Cell-Cycle Analysis Using Continuous Bromodeoxyuridine Labeling and Hoechst 33358-Ethidium Bromide Bivariate Flow Cytometry -- I. Introduction -- II. Application -- III. Materials -- IV. Procedures -- V. Controls and Standards -- VI. Instruments -- VII. Results -- References -- Chapter 22. Detection of BrdUrd-Labeled Cells by Differential Fluorescence Analysis of DNA Fluorochromes: Pulse-Chase and Continuous Labeling Methods -- I. Introduction -- 11. Applications -- 111. Materials -- IV. Cell Preparation and Staining Procedure -- V. Critical Aspects -- VI. Controls and Standards -- VII. Instruments -- VIII. Results -- References -- Chapter 23. Analysis of Intracellular Proteins -- I. Introduction -- II. Application -- III. Materials -- IV. Staining Procedures -- V. Critical Aspects -- VI. Controls and Standards -- VII. Instruments -- VIII. Multiparameter Analysis of Intracellular Antigens -- References -- Chapter 24. "Washless" Procedures for Nuclear Antigen Detection -- I. Introduction -- II. Applications -- Ill. Materials, Cell Preparation, and Staining. , IV. Critical Aspects of the Procedure: Controls, Standards, and Instruments -- V. Results -- References -- Chapter 25. Light Scatter of Isolated Cell Nuclei as a Parameter Discriminating the Cell-Cycle Subcompartments -- I. Introduction -- II. Application -- III. Materials -- IV. Critical Aspects of the Procedure -- V. Instruments -- Vl. Results -- VII. Appendix -- References -- Chapter 26. Simultaneous Analysis of Cellular RNA and DNA Content -- I. Introduction -- II. Applications -- III. Materials -- IV. Staining Procedures Employing A O -- V. Staining RNA and DNA with PY and Hoechst 33342 -- VI. Critical Aspects of the Procedures -- VII. Controls and Standards -- VIII. Instruments -- IX. Results -- X. Comparison of the Methods -- References -- Chapter 27. Analysis of DNA Content and Cyclin Protein Expression in Studies of DNA Ploidy, Growth Fraction, Lymphocyte Stimulation, and the Cell Cycle -- I. Introduction -- II. Bivariate Analysis of DNA Content versus Cyclin B1 or Cyclin E Expression -- III. Application of Cyclin B1 Analysis in DNA Ploidy and Chromatin Cycle Estimates -- IV. Lymphocyte Stimulation -- V. Tumor Growth Fraction Analysis -- Vl. Mapping the Cell Cycle: Mechanism of Action of Cytostatic Drugs -- VII. Critical Points of the Procedure -- VIII. Conclusions -- References -- Chapter 28. Oxidative Product Formation Analysis by Flow Cytometry -- I. Introduction -- II. Oxidative Burst -- III. Phagocytosis -- IV. Controls and Standards -- References -- Chapter 29. Flow Cytometric Determination of Cysteine and Serine Proteinase Activities in Living Cells with Rhodamine 110 Substrates -- I. Introduction -- II. Materials and Methods -- III. Results -- IV. Discussion -- References -- Chapter 30. Leucine Aminopeptidase Activity by Flow Cytometry -- I. Introduction -- II. Application -- III. Materials -- IV. Methods. , V. Critical Aspects of the Procedure.

Note: Front Cover -- Methods in Cell Biology -- CONTENTS -- Contents of Volume 64 -- Contributors -- Preface to the Third Edition -- Preface to the Second Edition -- Preface to the First Edition -- Part I: Principles of Cytometry and General Methods -- Chapter 1. A Brief History of Flow Cytometry and Sorting -- I. Introduction -- II. Instrumentation -- III. Applications -- References -- Chapter 2. Principles of Flow Cytometry: An Overview -- I. Introduction -- II. The Illumination of a Particle -- III. Fluidics: Centering Particles in the Illuminating Beam -- IV. Collection of Light Signals from Particles -- V. From Light Signals to a Data File -- VI. From Data to Information -- VII. Sorting -- VIII. Conclusions -- References -- Chapter 3. Laser Scanning Cytometry -- I. Introduction -- II. Background -- III. Description of the Instrument -- IV. The Utility and Operational Characteristics of Some Laser Scanning Cytometry List Mode Features -- V. Utility of Solid Phase Cytometry for Cell Preparation -- VI. Future Directions -- References -- Chapter 4. Principles of Confocal Microscopy -- I. Brief History of Microscope Development -- II. Development of Confocal Microscopy -- III. Image Formation in Confocal Microscopy -- IV. Useful Fluorescent Probes for Confocal Microscopy -- V. Applications of Confocal Microscopy -- VI. Conclusions -- References -- Chapter 5. Optical Measurements in Cytometry: Light Scattering, Extinction, Absorption, and Fluorescence -- I. Introduction -- II. Signal Processing Tasks in Flow Cytometry: An Overview -- III. The Optical Signal: Interaction of Light with Cells -- IV. Detection: Converting Optical Signals to Current -- V. Electronics: Converting Current to Voltage -- VI. Fluorescence Compensation and Logarithmic Amplification -- VII. Peak Detection, Integration, and Pulse Width Measurement. Triggering. , VIII. Measurement Sensitivity: Changing Concepts and the Bottom Line -- References -- Chapter 6. Flow Cytometric Fluorescence Lifetime Measurements -- I. Introduction -- II. Applications of the Technology -- III. Cell Preparation and Staining -- IV. Fluorescence Lifetime Flow Cytometry Instrumentation -- V. Results -- VI. Critical Aspects of the Technology -- VII. Future Directions -- References -- Chapter 7. Principles of Data Acquisition and Display -- I. Introduction -- II. Pulse Characterization Using Analog and Hybrid Circuits -- III. Analog Data Display -- IV. Analog-to-Digital Conversion -- V. Pulse Characterization by Digital Signal Processing -- VI. Data Storage and Display with Digital Computers -- References -- Chapter 8. Time as a Flow Cytometric Parameter -- I. Introduction -- II. Historical Overview -- III. Sample Mixing and Delivery -- IV. Data Analysis -- V. Applications -- VI. Conclusions -- References -- Chapter 9. Protein Labeling with Fluorescent Probes -- I. Introduction -- II. Labeling of Proteins with Organic Fluorescent Dyes -- III. Labeling of Proteins with Phycobiliproteins -- IV. Conclusion -- References -- Part II: Cell Preparation -- Chapter 10. Preparation of Cells from Blood -- I. Introduction -- II. Collection, Transport, and Storage of Blood -- III. Fixation and Preservation -- IV. Separation of Erythrocytes from Leukocytes -- V. Assessment of Cell Viability -- VI. Staining -- VII. Summary -- Appendix 1 -- Appendix 2 -- Appendix 3 -- References -- Chapter 11. Cell Preparation for the Identification of Leukocytes -- I. Introduction -- II. Antibodies -- III. Tandem Fluorochromes -- IV. Cell Preparation and Staining Procedures -- V. Titering Antibodies -- VI. Solutions and Reagents -- References -- Chapter 12. Strategies for Cell Permeabilization and Fixation in Detecting Surface and Intracellular Antigens. , I. Introduction -- II. Application -- III. Materials and Methods -- IV. Concluding Remarks -- References -- Part III: Standardization, Quality Assurance -- Chapter 13. Stoichiometry of Immunocytochemical Staining Reactions -- I. Introduction -- II. Structure of Immunoglobulin G -- III. Cell Structure -- IV. Permeabilized Cell Structure -- V. Antibody-Antigen Reactions -- VI. Multiparametric Analyses -- VII. Summary -- References -- Chapter 14. Standardization and Quantitation in Flow Cytometry -- I. Introduction -- II. General Issues -- III. Performance Characteristics-Dynamic Range, Linearity, Resolution, and Sensitivity -- IV. Standardization and Calibration of Common Cytometry Measurements -- V. Examples of Applications Using Calibrated Measurements -- VI. Issues in Quantitation of Fluorochromes and Other Molecules -- References -- Part IV: Cell Proliferation -- Chapter 15. Methods to Identify Mitotic Cells by Flow Cytometry -- I. Introduction -- II. Materials -- III. Cell Preparation and Staining -- IV. Instruments -- V. Critical Aspects of the Procedure -- VI. Results and Discussion -- VII. Comparison of Anti-H3-P Monoclonal Antibody with Other Markers of Mitotic Cells -- References -- Chapter 16. Cell Cycle Kinetics Estimated by Analysis of Bromodeoxyuridine Incorporation -- I. Introduction -- II. Applications -- III. Materials -- IV. Methods -- V. Critical Aspects of the Procedure -- References -- Chapter 17. Flow Cytometric Analysis of Cell Division History Using Dilution of Carboxyfluorescein Diacetate Succinimidyl Ester, a Stably Integrated Fluorescent Probe -- I. Introduction and Background -- II. Reagents and Solutions -- III. Preparation and Labeling of Cells -- IV. Gathering of Information Concurrent with Division -- V. Analysis of Data. , VI. Application of Carboxyfluorescein Diacetate Succinimidyl Ester to in Vitro Culture of Lymphocytes -- VII. Monitoring Lymphocyte Responses in Vivo -- VIII. Antigen Receptor Transgenic Models -- References -- Chapter 18. Antibodies against the Ki-67 Protein: Assessment of the Growth Fraction and Tools for Cell Cycle Analysis -- I. Introduction -- II. Application -- III. Materials and Methods -- IV. Critical Aspects -- V. Controls and Standards -- VI. Examples of Results -- References -- Chapter 19. Detection of Proliferating Cell Nuclear Antigen -- I. Introduction -- II. Molecular Biology of Proliferating Cell Nuclear Antigen -- III. Methods for Immunochemical Detection and Quantification of Proliferating Cell Nuclear Antigen -- IV. Results of Cytometric Analysis of Proliferating Cell Nuclear Antigen Expression -- V. Applications in Toxicology, Pathology, and Oncology -- References -- Chapter 20. Lymphocyte Activation Associated Antigens -- I. Introduction -- II. Methodological Aspects -- III. To Flow or Not to Flow for Assessing Lymphocyte Activation/ Proliferation? And, If Yes, How Reliable Is Immunophenotyping? -- IV. Additional Approaches -- V. Concluding Remarks -- References -- Part V: Cell Death/Apoptosis -- Chapter 21. Analysis of Mitochondria during Cell Death -- I. Introduction -- II. Scientific Background -- III. Apoptosis and Mitochondria -- IV. Method -- V. Results -- VI. Pitfalls and Misinterpretation of the Data -- VII. Comparison with Other Methods -- VIII. Reviews of the Applications -- IX. Biological and Biomedical Information -- X. Future Directions -- References -- Chapter 22. Cytometry of Caspases -- I. Introduction -- II. Materials and Methods: Caspase Peptide Inhibitors and Methods to Monitor Responses -- III. Results and Discussion -- References -- Chapter 23. Analysis of Apoptosis in Plant Cells -- I. Introduction. , II. Apoptosis in Plants -- III. Problems Associated with Analyzing Plant Cells Using Flow Cytometry -- IV. Morphological Changes of Plant Cells -- V. Physiological Changes during Apoptosis -- VI. Conclusion -- References -- Chapter 24. Difficulties and Pitfalls in Analysis of Apoptosis -- I. Introduction -- II. Apoptotic Index May Not Be Correlated with Incidence of Cell Death -- III. Dif.culties in Estimating Frequency of Apoptosis by Analysis of DNA Fragmentation -- IV. The Lack of Evidence Is Not Evidence for the Lack of Apoptosis -- V. Misclassification of Apoptotic Bodies or Nuclear Fragments as Single Apoptotic Cells -- VI. Apoptosis versus Necrosis versus ''Necrotic Stage'' of Apoptosis -- VII. Selective Loss of Apoptotic Cells during Sample Preparation -- VIII. Live Cells Engulfing Apoptotic Bodies Masquerade as Apoptotic Cells -- IX. The Problems with Commercial Kits and Reagents -- X. Cell Morphology Is Still the Gold Standard for Identification of Apoptotic Cells -- XI. Laser Scanning Cytometry: Have Your Cake and Eat It Too -- References -- Part VI: Cell-Cell, Cell-Environment Interactions -- Chapter 25. Analysis of Cell Migration -- I. Introduction and Application -- II. General Strategies to Measure Cell Migration -- References -- Chapter 26. Three-Dimensional Extracellular Matrix Substrates for Cell Culture -- I. Introduction -- II. Application -- III. Methods -- IV. Application of Intestinal Submucosa as a Three-Dimensional Extracellular Matrix Substrate -- V. Summary -- References -- Chapter 27. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix-Cell Interactions -- I. Introduction -- II. Three-Dimensional Imaging of Extracellular Matrix and Extracellular Matrix-Cell Interactions: Current Techniques and Their Limitations. , III. Three-Dimensional Microscopy of Living Systems: Extracellular Matrix and Extracellular Matrix-Cell Interactions.

Note: Front Cover -- Methods in Cell Biology -- Copyright Page -- Contents -- Contributors -- Preface -- Chapter I. Biophysics of the Green Fluorescent Protein -- I. Introduction -- II. Protein Folding and the Generation of This Chromophore -- III. The Biophysics of the Fluorescence of GTP -- IV. Resonance Energy Transfer Involving GFP -- V. Summary -- References -- Chapter 2. Understanding Structure-Function Relationships in the Aequorea Victoria Green Fluorescent Protein -- I. Introduction -- II. Structure -- III. Chromophore Formation -- IV. Effects of Mutations on the Spectroscopic Properties of GFP -- V. Effects of Mutations That Improve Thermosensitivity -- VI. The Development of Enhanced Mutants -- References -- Chapter 3. Quantitative Imaging of the Green Fluorescent Protein (GFP) -- I. Introduction -- II. Factors That Influence/Limit Quantitation of GFP in Fluorescence Microscopy -- III. Applications of LSCM for Quantitative Imaging of GFP -- IV. Preparation of Purified GFP Samples -- References -- Chapter 4. Single-Molecule Fluorescence Detection of Green Fluorescence Protein and Application to Single-Protein Dynamics -- I. Introduction -- II. Design Considerations for Fluorescence Microscopes for Single-Molecule Detection -- III. Characteristics of the Fluorescence from Single GFP Molecules -- IV. Advantages of Using GFP for Single-Molecule Detection -- V. GFP in Vitro and in Vivo Assays -- Appendix I. Details of the TIR Microscope -- Appendix II. Data Acquisition and Analysis -- References -- Chapter 5. Targeting GFP to Organelles -- I. Introduction -- II. Construction and Expression of the Organelle-Targeted GFP Chimeras -- III. Dynamic Monitoring of Organelle Structure with the Targeted GFPs -- IV. Expression in Primary Cultures -- V. Visualizing GFP Chimeras with Different Spectral Properties -- VI. Protocols -- References. , Chapter 6. Cytoskeletal Dynamics in Yeast -- I. Introduction -- II. Generating GFP Fusions -- III. Imaging Considerations for Yeast Cells -- IV. Time-Lapse Microscopy -- V. Results: Cytoskeletal GFP Fusion Proteins -- VI. Future -- References -- Chapter 7. Analysis of Nuclear Transport in Vivo -- I. Introduction -- II. Experimental Approaches and Protocols -- References -- Chapter 8. GFP Fusion Proteins as Probes for Cytology in Fission Yeast -- I. Introduction -- II. Expressing GFP Fusion Proteins -- III. Applications of Fusion Proteins -- References -- Chapter 9. GFP Variants for Multispectral Imaging of Living Cells -- 1. Introduction -- II. Green Fluorescent Protein Markers -- III. Imaging of Living Cells -- IV. Marking Different Cell Types in Arabidopsis -- V. Spectrally Distinct Fluorescent Proteins for Multichannel Confocal Microscopy -- VI. Summary -- References -- Chapter 10. GFP Fusions to a Microtubule Motor Protein to Visualize Meiotic and Mitotic Spindle Dynamics in Drosophila -- I. Introduction -- II. Labeling Strategies -- III. Imaging GFP -- IV. Applications of Ncd-GFP Imaging -- V. Perspectives -- References -- Chapter 11. GFP as a Cell and Developmental Marker i n the Drosophila Nervous System -- I. Introduction -- II. Targeted Expression of GFP in Drosophila -- III. Lines for Expression of GFP -- IV. Visualizing GFP Expression -- References -- Chapter 12. Using Time-Lapse Confocal Microscopy for Analysis of Centromere Dynamics in Human Cells -- I. Introduction -- II. GFP Fusion Proteins -- III. Microscopy -- IV. Analysis -- V. Summary -- Appendix: Handling Confocal Images on the Laboratory Computer -- References -- Chapter 13. Visualization of Largescale Chromatin Structure and Dynamics Using the lac Operator/lac Repressor Reporter System -- I. Introduction -- II. Overview of Methodology. , III. Construction of the lac Operator Repeat -- IV. Manipulation of the lac Operator Repeats -- V. Repressor-NLS and GFP-Repressor-NLS Constructs -- VI. Gene Amplification and Cell Cloning -- VII. Repressor Staining and Immunodetection of the lac Operator Repeat -- VIII. In Vivo Observation of GFP-Repressor Localization -- IX. Phototoxicity Issues -- X. Present Results and Future Directions -- References -- Chapter 14. Centrosome Dynamics in Living Cells -- I. Introduction -- II. Cloning and Expression of GFP-Pericentrin -- III. High-speed Microscopy -- IV. Image Restoration by an Improved Deconvolution Method -- V. Imaging Centrosomes -- VI. Postimaging Confirmation of Centrosome Integrity and Function -- References -- Chapter 15. Transfections of Primary Muscle Cell Cultures with Plasmids Coding for GFP Linked to Full-Length and Truncated Muscle Proteins -- I. Introduction -- II. Construction of GFP-Linked Muscle Proteins -- III. Preparation of Embryonic Avian Cardiomyocytes and Skeletal Muscle Myoblasts -- IV. Methods of Transfection of Cross-Striated Cells in Culture -- V. Transfection of Cross-Striated Muscle Cells with Full-Length cDNA for Sarcomeric Proteins -- VI. Microscopic Observations of Live Cells -- VII. Postprocessing of Transfected Cells -- VIII. Problems Encountered in Cells Transfected with GFP-Sarcomeric Proteins -- IX. Overview -- References -- Chapter 16. Monitoring the Dynamics and Mobility of Membrane Proteins Tagged with Green Fluorescent Protein -- I. Introduction -- II. Constructing and Expressing GFP Fusion Proteins: Strategies for Optimizing Brightness and Assessing Chimera Function -- III. Practical Guidelines for the Preparation and Imaging of GFP-Expressing Cells -- IV. Time-Lapse Imaging of GFP Chimeras: Critical Parameters -- V. Analysis of Time-Lapse Imaging Data. , VI. Relating GFP Chimera Fluorescence to Actual Numbers of GFP Molecules -- VII. Fluorescence Recovery after Photobleaching FRAP -- VIII. Qualitative FRAP Experiments -- IX. Quantitative FRAP -- X. Calculating D -- XI. Fluorescence Loss in Photobleaching (FLIP) Using a Confocal Microscope -- XII. Other Applications of Photobleaching -- References -- Chapter 17. Synchronous Real-Time Reporting of Multiple Cellular Events -- I. Introduction -- II. Green Fluorescent Protein -- III. Luciferase -- IV. Instrumentation and Techniques -- V. Future Directions -- References -- Chapter 18. Visualizing Protein Interactions in Living Cells Using Digitized GFP Imaging and FRET Microscopy -- I. Introduction -- II. The Theory of FRET -- III. Review of the FRET Literature -- IV. Why Use FRET Microscopy? -- V. Why Use the GFPs for FRET? -- VI. Some Considerations for the Use of GFPs in FRET Imaging -- VII. Some Considerations for Designing a FRET Imaging System -- VIII. The Practical Application of FRET to Visualize Protein- Protein Interactions -- IX. Overview and Conclusion -- References -- Chapter 19. Flow Cytometric Analysis and FACS Sorting of Cells Based on GFP Accumulation -- I. General Introduction -- II. Methods and Specific Applications -- III. Typical Results -- IV. Discussion and Conclusions -- References -- Chapter 20. GFP Biofluorescence: Imaging Gene Expression and Protein Dynamics in Living Cells -- I. Introduction -- II. Facilities -- III. Maintaining Cells -- IV. Imaging System -- V. Computer Systems -- VI. output -- VII. Conclusions -- References -- Index.

Note: Front Cover -- Microbes as Tools for Cell Biology -- Copyright Page -- Contents -- Contributors -- Preface -- Introduction -- Chapter 1. Obtaining and Maintaining Microbial Pathogens -- I. Introduction -- II. Biosafety Considerations -- III. Department of Health and Human Services Guidelines -- IV. Federal and Institutional Requirements -- V. Source of Pathogens -- VI. Culture and Maintenance of Pathogens -- VII. Conclusion -- Part I: Manipulation of Pathogens -- Chapter 2. Cultivation of Malaria Parasites -- I. Introduction -- II. Culture Medium -- III. Serum -- IV. Erythrocytes -- V. Culture Systems -- VI. Synchronization -- VII. Gametocyte Production in Culture -- VIII. Cloning Methods -- IX. Cryopreservation -- X. Serum Replacement -- XI. Axenic (Extracellular) Development of Erythrocytic Stages of P. falciparum -- XII. Cultivation of Erythrocytic Stages of Other Species of Malaria Parasites -- XIII. In Vitro Development of Preerythrocytic and Sporogonic Stages of the Life Cycle -- References -- Chapter 3. Molecular Tools for Genetic Dissection of the Protozoan Parasite Toxoplasma gondii -- I. Introductory Overview -- II. In Vitro Culture of T. gondii Tachyzoites -- III. Molecular Transformation Systems for Toxoplasma -- IV. Summary and Outlook -- References -- Chapter 4. Transfection Experiments with Leishmania -- I. Introduction -- II. Stable and Transient Transfection -- III. Transfection -- References -- Chapter 5. Mutagenesis and Variant Selection in Salmonella -- I. Introduction -- II. Mutagenesis -- III. Screening of Variants -- IV. Genetic Analysis -- V. Perspectives -- References -- Chapter 6. Mycobacterium: Isolation, Maintenance, Transformation, and Mutant Selection -- I. Introduction -- II. Biosafety Considerations -- III. Culture Media and Conditions -- IV. Isolation of Mycobacteria -- V. Maintenance of Stocks. , VI. Genetic Techniques -- VII. Mutagenesis -- VIII. Strain Construction -- IX. Mutant Selection and Isolation -- References -- Part II: Microbial Adherence and Invasion Assays -- Chapter 7. Modulation of Murine Macrophage Behavior in Vivo and in Vitro -- I. Introduction -- II. Reagents and Solutions -- III. Animal Husbandry and Maintenance of Immunocompromised Mice -- IV. Eliciting and Harvesting Macrophages in Vivo -- V. Adherence of Macrophages to Solid Substrates -- VI. Assays for Macrophage Phagocytic Activity -- VII. Analysis of MHC Class II Antigen Expression -- References -- Chapter 8. In Vitro Assays of Phagocytic Function of Human Peripheral Blood Leukocytes: Receptor Modulation and Signal Transduction -- I. Introduction -- II. Phagocytosis Assays -- III. Phagocytic Receptors -- IV. Stimulation of Phagocytosis -- V. Phagosome Isolation -- References -- Chapter 9. Bacterial Adhesion and Colonization Assays -- I. Introduction -- II. In Situ Screening of Host Receptor Distribution -- III. Bacterial Adherence to Cells in Culture -- IV. Biochemical Characterization of the Molecular Nature of Receptors in Situ -- V. Bacterial Inhibition Experiments in Situ -- VI. In Vitro Assays for Bacterial Adhesion -- VII. Probing Eukaryotic Cell Glycoconjugates with Purified Bacterial Adhesins -- VIII. Concluding Remarks -- References -- Chapter 10. Cytoadherence and the Plasmodium falciparum-Infected Erythrocyte -- I. Introduction -- II. Ligands for Adherence -- III. P. falkiparum-Infected Red Cell Adhesions -- IV. Cytoadherence, an in Vitro Model of Sequestration: Practical Considerations of Cytoadherence Assays -- References -- Part III: The Study of Intracellular Pathogenesis -- Chapter 11. Purification of Plasmodium falciparum Merozoites for Analysis of the Processing of Merozoite Surface Protein-1 -- I. Introduction. , II. Parasite Culture and Synchronization -- III. Merozoite Isolation -- IV. Assay for Secondary Processing of the Merozoite Surface Protein-1 (MSP-1) -- References -- Chapter 12. In Vitro Secretory Assays with Erythrocyte-Free Malaria Parasites -- I. Inroduction -- II. Release and Separation of Late Ring and Trophozoite Stage Parasites from the Erythrocyte Membrane (EM) and Tubovcsicular -- III. Synthesis and Secretion of Proteins by Intact, Ring/Trophozoite Parasites -- IV. Organization of Secretory Activities at Different Stages of the Asexual Life Cycle -- V. Release of Pigmented Trophozoits and Schizonts from Infected Erythrocytes by Osmotic Shock in Isoosmolar Dipeptide-Based Medid -- References -- Chapter 13. Intracellular Survival by Legionella -- I. Introduction -- II. Laboratory Cultivation of L. pneumophila -- III. Tissue Culture of U937 Cell-Derived Macrophages -- IV. Intracellular Thymineless Death Enrichment -- V. Identification of Intracellular Growth Mutants from Enriched Bacterial Pools Using "Poke Plaque" Assays -- VI. Additional Remarks -- References -- Chapter 14. Isolation and Characterization of Pathogen-Containing Phagosomes -- I. Introduction -- II. Choice of Pathogens and Particles -- III. Choice of Macrophage -- IV. Particle Adherence and Internalization Conditions -- V. Cell Lysis Conditions -- VI. Isolation of Phagosomes -- VII. Analysis of Phagosomal Constituents -- VIII. Storage and Handling of Two-Dimensional SDS-PAGE Data -- IX. Shortcomings -- References -- Chapter 15. Immunoelectron Microscopy of Endosomal Trafficking in Macrophages Infected with Microbial Pathogens -- I. Introduction -- II. The Host-Pathogen Interplay -- III. Intersection with the Endosomal Pathway -- IV. Processing of Infected Macrophages for Immunoelectron Microscopy -- V. Blocking Cryosections and Incubation with Primary Antiserum. , VI. Gold-Conjugated Second Antibodies. -- VII. Controls -- VIII. Final Preparation of the Grids -- IX. Routine Protocol for Analysis of Fluid-Phase Trafficking -- References -- Chapter 16. Measuring the pH of Pathogen-Containing Phagosomes -- I. Introduction -- II. Materials -- III. Procedures -- IV. When Things Are Not Perfect, or Even Very Close -- V. Conclusion -- References -- Chapter 17. Techniques for Studying Phagocytic Processing of Bacteria for Class I or II MHC-Restricted Antigen Recognition by -- I. Introduction -- II. Generating MHC-I- and MHC-II- Restricted T Cells to Detect Antigen Processing -- III. Antigen Presentation Assays -- IV. Observations and Implications -- References -- Index -- Volumes in Series.

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.