Note: Front Cover -- Methods in Cell Biology, Volume 27 -- Copyright Page -- Contents -- Contributors -- Preface -- Chapter 1. Laboratory Culture of Strongylocentrotus purpuratus Adults, Embryos, and Larvae -- I. Introduction -- II. Methods for Management of Adult Sea Urchins -- III. Embryo Culture and Larval Growth -- IV. Metamorphosis and Maintenance of Juveniles -- References -- Chapter 2. Handling, Labeling, and Fractionating Sea Urchin Spermatozoa -- I. Introduction -- II. Seasonality, Quantity, and Methods of Collection -- Ill. Artificial Seawaters -- IV. Preparing a Pure Sperm Suspension -- V. The Effect of Storage on Respiration and Fertilizability -- VI. Determining Sperm Concentration and Quantitating Fertilizability -- VII. Detecting and Quantitating the Acrosome Reaction -- VIII. Labeling Sperm Components -- IX. Isolation of Sperm Components -- References -- Chapter 3. Sperm Motility -- I. Experimental Preparation -- II. Observation and Measurement -- References -- Chapter 4. Ion Measurements in Sea Urchin Sperm -- I. Introduction -- II. Sperm Preparation and Determination of Intracellular Water Space -- III. pH Measurements Important to Sperm Activation -- IV. Measurement of Cation Uptake -- V. Measurement of Membrane Potentials in Sperm -- VI . Conclusion -- References -- Chapter 5. Gonad-Stimulating and Maturation-Inducing Substance -- I. Introduction -- II. Gonad-Stimulating Substance -- III. Maturation-Inducing Substance -- IV. Outlook -- References -- Chapter 6. Techniques for Observing Living Gametes and Embryos -- I. Introduction -- II. Handling and Care of Animals -- III. Obtaining Gametes -- IV. "Biocleaning" Glassware -- V. Fertilization and Culturing -- VI. Observation Chambers -- VII. Considerations for Microscopic Observations of Development of Single Embryos. , VIII. Iontophoretic Injection of Lucifer Yellow CH to Study Embryonic Cell Lineage -- References -- Chapter 7. Egg and Embryonic Extracellular Coats: Isolation and Purification -- I. Preparation of Gametes -- II. Isolation of Fertilization Membrane Precursors -- III. Fertilization and Parthenogenetic Activation of Eggs -- IV. Isolation of Fertilization Membrane Intermediates -- V. Isolation of Soluble Fertilization Product -- VI. Methods for Concentrating Fertilization Product -- VII. Purification of Components from Fertilization Product -- VIII. Techniques for the Analysis of Fertilization Membrane Proteins -- References -- Chapter 8. Purification and Properties of the Egg Plasma Membrune -- I. Introduction -- II. Plasma Membranes from Unfertilized Sea Urchin Eggs -- III. Plasma Membranes from Fertilized Eggs -- IV. Characterization of Egg Plasma Membranes -- References -- Chapter 9. Nuclei and Chromosomal Proteins -- I. Introduction -- II. Inducing Polyspermy -- III. Cell Cycle Parameters -- IV. Isolation of Nuclei -- V. Protein Analysis -- VI. Conclusions -- References -- Chapter 10. The Protein Synthetic Machinery: Ribosomes and Cell-Free Systems -- I. Introduction -- II. Strategy -- III. The Isolation of Ribosomes and Polyribosomes -- IV. A Ribosome-Free Translation System for Testing Echinoid Ribosomes -- V. Isolation of Inhibitory Proteins -- VI. Homologous Cell-Free Echinoid Translational Systems -- VII. Concluding Remarks -- References -- Chapter 11. Tubulin-Containing Structures -- I. Introduction -- II. Unfertilized Egg and Oocyte Tubulin -- III. Mitotic Spindle Tubulin -- IV. Cilia and Flagella -- V. Properties of Microtubules Reassembled from Egg, Spindle, and Flagellar Tubulins -- VI. Microtubule-Associated Proteins -- VII. Mitotic Organizing Centers -- VIII. Particulate Tubulin -- IX. Concluding Remarks -- References. , Chapter 12. Isolation of Embryonic Cilia and Sperm Flagella -- I. Introduction -- II. Isolation Methods -- III. Fractionation Methods -- References -- Chapter 13. Components of the Actin-Based Cytoskeleton -- I. Introduction -- II. Preparation of Extracts -- III. Behavior of Extracts and Preparation of Cytoskeletal Proteins -- IV. Problems and Prospects -- References -- Chapter 14. Cytology and Immunocytochemistry -- I. Introduction -- II. Whole Mounts of Fixed and Embedded Material -- III. Sectioned and Stained Material for Bright-Field Light Microscopy -- IV. Indirect Immunofluorescence Microscopy -- V. Other Fluorochrome Probes -- References -- Chapter 15. Scanning Electron Microscopy of Embryos -- I. Introduction -- II. Specimen Preparation Supplies -- III. Specimen Preparation -- IV. Artifacts of Critical Point-Dried Specimens -- V. Operating Parameters and Secondary Electron Imaging with the Scanning Electron Microscope -- VI. Examining and Photographing Embryos -- VII. Recording Stereo Pair Images -- VIII. Photomontages -- IX . SEM of Sectioned Embryos -- X. Photoetching and Line Drawings -- XI. Conclusions -- References -- Chapter 16. Cytological Techniques for the Study of Larval Echinoids with Notes on Methods for Inducing Metamorphosis -- I. Introduction -- II. Monoclonal Antibodies against Tissue-Specific Antigens in Sea Urchin Embryos -- III. Visualization of Larval Nerves with Glyoxilic Acid-Induced Fluorescence -- IV. Induction of Metamorphosis in Echinoid Larvae -- References -- Chapter 17. Embryo Dissociation, Cell Isolation, and Cell Reassociation -- I. Introduction -- II. Embryo Dissociation -- III. Gem Layer Separation -- IV. Reassociation Methods -- V. Antibodies and Measurement of Cell Interactions -- VI. Summary -- References -- Chapter 18. Manipulative Methods for Analyzing Embryogenesis -- I. Introduction. , II. Stripping Eggs -- III. Bisection of Sea Urchin and Starfish Eggs -- IV. Formation of Fused Starfish Embryos -- V. Marking Starfish Blastomeres with Injected Horseradish Peroxidase -- VI. Chromosome Staining -- References -- Chapter 19. Simple Methods and Devices for Handling Echinoderm Eggs -- I. Introduction -- II. Equipment for Making Instruments -- III. Fabrication and Care of Instruments -- IV. Instruments and Devices -- V. Manipulations -- VI. Summary -- References -- Chapter 20. Induced Fusion of Echinoderm Oocytes and Eggs -- I. Introduction -- II. Fusion between Starfish Oocytes and between Oocytes and Cytoplasts -- III. Fusion of Mature Unfertilized Sea Urchin Eggs -- IV. Concluding Remarks -- V. Appendix: Methods for Maintaining Starfish and Obtaining Gametes and Embryos Year Round -- References -- Chapter 21. Microinjection and Cytoplasmic Transfer in Starfish Oocytes -- I. Introduction -- II. Apparatus for Microinjection -- III. Procedures for Microinjection -- IV. Application of Microinjection Techniques in Starfish to the Study of Oocyte Maturation -- V. Concluding Remarks -- References -- Chapter 22. Rapid Repetitive Microinjection -- I. Introduction -- II. General Method -- III. Preparations -- IV. Injecting the Cells -- V. Results of Microinjection and Gel Electrophoresis of Proteins -- VI. Summary -- References -- Chapter 23. Electrophoresis of Eggs and Blastomeres -- I. Introduction -- II. Theory -- III. Apparatus -- IV. Measurement of Electrophoretic Mobility -- V. Application -- References -- Chapter 24. The Compression Method for Determining the Surface Force -- I. Introduction -- II. The Bending Balance as a Monitor of the Force of Compression -- III. Experimental Set-Up -- IV. Practicals of Measurements -- V. Calculation of the Surface Force -- VI. Weight Loading Method -- VII. Critical Comments on the Methods. , References -- Chapter 25. Determination of Mechanical Properties of the Egg Surface by Elastimetry -- I. Introduction -- II. Principles of the Elastimetry -- III. Experimental Set-Up -- IV. Practicals of Measurement -- V. Mechanical Properties of the Cell Surface -- References -- Chapter 26. Determination of Mechanical Properties of the Egg by the Sessile Drop Method -- I. Introduction -- II. Measurement of the Form of the Egg -- III. Determination of Mechanical Properties of the Egg -- IV. Limitations of the Present Method and Errors in Measurement -- V. Appendix: Derivation of the Profiles of Sessile Drops -- References -- Index.

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