Collected Works Of Shinya Inoué: Microscopes, Living Cells, and Dynamic Molecules (With DVD-ROM

This book collects the publications of Shinya lnoué, pioneering cell biophysicist and winner of the 2003 International Prize for Biology. The articles cover the discovery, and elucidate the behavior in living cells, of the dynamic molecular filaments which organize the cell and play a central role in cell division. Other articles report on the development of microscopes, especially those using polarized light and digital image enhancement, which make possible studies of the ever-changing molecular architecture directly in living cells. This book also contains many high quality photo-micrographs as well as an appended DVD with an extensive collection of video movies of active living cells. After training in Tokyo and at Princeton University, Dr Inoué has held teaching positions at the University of Washington, Tokyo Metropolitan University, University of Rochester, Dartmouth Medical School, and University of Pennsylvania. He is a member of the U.S. National Academy of Sciences and currently holds the title of Distinguished Scientist at the Marine Biological Laboratory in Woods Hole, Massachusetts. Contents: Introduction to Doctoral Thesis; Physical Chemistry of Microtubules in vivo; Axostyle Motility in Pyrsonympha; Cell Division and the Mitotic Spindle -- A Review; Recollection of Kayo Okazaki; Foundations of Confocal Microscopy; Centrifuge Polarizing Microscope: Design, Performance and Biological Applications; Theory, Measurements and Rectification of Polarization Aberrations; Fluorescence Anisotropy of GFP Crystals; Biological Polarization Microscopy; Address by Emperor Akihito and Acceptance Speech by Shinya Inoué: International Prize for Biology; and other papers. CONTENTS......Page 6 Introduction and Guide......Page 12 Using the DVD with a Windows Computer......Page 14 Viewing the Slides and Movies in PowerPoint......Page 15 Hyperlinks to Additional Material......Page 16 Article 1 Compressorium design (in Botany & Zoology, 1943)......Page 24 2.1. Detailed explanation of the invention......Page 26 2.2. Patent claims......Page 28 Article 3 Birefringence vs. length and contraction of single muscle fiber (unpublished, 1947)......Page 30 References......Page 33 Article 4 Cover slip thickness gauge (unpublished, 1951)......Page 34 References......Page 36 Article 5 Birefringence of the dividing cell (with Dan K, in J Morphol, 1951) with note added in 2006......Page 38 METHODS......Page 39 RESULTS......Page 44 DISCUSSION......Page 58 SUMMARY......Page 60 APPENDIX......Page 61 REFERENCES......Page 65 PLATE 1......Page 67 PLATE 2......Page 69 Article 6 Introduction to doctoral thesis (unpublished, 2006)......Page 72 ACKNOWLEDGMENTS......Page 74 1. Introduction......Page 76 References......Page 80 Article 8 Thesis Part II. Depolarization of light by microscope optics (in Exp Cell Res, 1952)......Page 82 CALCULATION OF THE ROTATION OF RAYS PASSING THROUGH DIFFERENT POINTS OF THE EXIT PUPIL......Page 84 THE TOTAL FLUX OF LIGHT INTRODUCED BY ROTATION WITHIN A GIVEN APERTURE......Page 89 REFERENCES......Page 91 Article 9 Thesis Part III. Device for measuring retardation in small objects (in Exp Cell Res, 1951)......Page 92 REFERENCES......Page 96 Introduction.......Page 98 1. The Structure of the spindle in the oocyte of Chaetopterus pergamentaceous.......Page 99 2. The spindle in the pollen mother cell of Lilium longiflorum.......Page 105 Literature.......Page 110 MATERIALS AND METHODS......Page 112 OBSERVATIONS ON THE EFFECT OF COLCHICINE......Page 114 DISCUSSION......Page 118 DISCUSSION......Page 121 1. The Submicroscopic Structure of the Spindle in Living Cells......Page 126 References......Page 129 The following note was added by Shinya Inoue in September of 2006:......Page 132 I. The Value and Limitation of Polarization Microscopy in Biology:......Page 134 II. The Polarization Rectifier:......Page 135 ///. The Polarizing Microscope After Rectification:......Page 140 EXPLANATION OF PLATES......Page 141 Article 15 Diffraction anomaly in polarizing microscopes (with Kubota H, in Nature, 1958)......Page 148 1. STRUCTURE AND MOTILITY OF CILIA......Page 152 2. MICROSCOPIC STRUCTURE OF THE MITOTIC SPINDLE......Page 153 3. CENTERS OF ORGANIZATION IN SPINDLE AND CILIA......Page 154 4. ON THE PHYSICOCHEMICAL NATURE OF THE MITOTIC SPINDLE......Page 155 BIBLIOGRAPHY......Page 157 2. DETRACTION IMAGE......Page 160 3. REFRACTING OBJECTIVE......Page 161 4. CATADIOPTRIC SYSTEM......Page 164 5. z-CTJT UNIAXIAL CRYSTAL......Page 165 7. ACKNOWLEDGMENTS......Page 167 General Description......Page 168 Specification of the Components......Page 169 REFERENCES......Page 172 Introduction......Page 174 Technique......Page 175 Observations......Page 177 Discussion and conclusion......Page 181 References......Page 188 Article 20 Heavy water enhancement of spindle birefringence (with Sato H and Tucker RW, in Biol Bull, 1963)......Page 190 Article 21 Rapid exchange of D2O and H2O in sea urchin eggs (with Tucker RW, in Biol Bull, 1963)......Page 192 Introduction......Page 194 Description of Spindle Birefringence......Page 197 Changes in Birefringence during Mitosis......Page 201 Motion Picture of Living Cells in Division......Page 215 Effect of Low Temperature4......Page 218 Thermodynamic Studies......Page 219 Orienting Centers—Ultraviolet Microbeam Experiments5......Page 225 Conclusion......Page 228 REFERENCES......Page 237 DISCUSSION......Page 239 Article 23 Heavy water counteracts effect of Colcemid on spindle (with Sato H and Ascher M, in Biol Bull, 1965)......Page 244 Birefringence of the Sperm Head......Page 246 Effect of Polarized Ultraviolet Irradiation......Page 247 Response of Sperm to Irradiation by UV Ray Polarized at 45°......Page 249 Photographic Microdensitometry......Page 250 Suggested Model......Page 252 Concluding Remarks......Page 253 REFERENCES......Page 254 Article 25 Cell motility by labile association of molecules (with Sato H, in J Gen Phys, 1967) with note added in 2006......Page 286 1. VISIBLE CHANGES OF SPINDLE FIBER BIREFRINGENCE DURING MITOSIS......Page 288 2. MECHANICALINTE GRITY OF SPINDLE FIBERS......Page 292 3. DYNAMIC NATURE OF SPINDLE FIBERS......Page 293 4. THE DYNAMIC EQUILIBRIUM MODEL......Page 294 5. EFFECT OF HEAVY WATER ON SPINDLE BIREFRINGENCE......Page 295 6. ASSEMBLY FROM POOL, NOT DE NOVO SYNTHESIS......Page 297 7. CONTROL OF ORIENTATION......Page 300 8. EXTENSION AND CONTRACTION OF SPINDLE FIBERS......Page 304 10. RELATION OF SPINDLE FIBER B I R E F R I N G E N C E TO MICROTUBULES AND TO FILAMENTS OBSERVED BY ELECTRON MICROSCOPY......Page 305 11. CELL MOTILITY AND LABILE ASSOCIATION OF MOLECULES......Page 308 REFERENCES......Page 309 Discussion......Page 315 RESULTS......Page 322 SUMMARY......Page 328 BIBLIOGRAPHY......Page 329 INTRODUCTION......Page 330 TEMPERATURE CONTROL AND RETARDATION RECORDING SYSTEM......Page 331 PRESSURE INDUCED CHANGE OF EQUILIBRIUM......Page 341 KINETICS OF SPINDLE RETARDATION CHANGE......Page 343 DISCUSSION......Page 347 REFERENCES......Page 348 ABSTRACT......Page 350 Imbibition Procedure......Page 351 Retardation Measurements......Page 354 Electron Microscopy......Page 355 Results of Imbibition......Page 358 FORM BR IN OTHER MATERIAL......Page 360 Form BR of Skeletal Muscle......Page 361 DISCUSSION......Page 362 REFERENCES......Page 365 Article 29 Crystal property of spicules in sea urchin pluteus (with Okazaki K, in Dev Growth Differ, 1976)......Page 368 MATERIALS AND METHODS......Page 369 RESULTS......Page 370 DISCUSSION......Page 381 REFERENCES......Page 388 ABSTRACT......Page 390 Culture of the Protozoa......Page 391 Development of Astral Rays and Central Spindle......Page 393 DISCUSSION......Page 398 REFERENCES......Page 405 ABSTRACT......Page 408 RESULTS......Page 409 Changing Contour of the Nucleus in Lateral View......Page 418 Anterior View and Three-Dimensional Interpretation of Nuclear Morphogenesis......Page 419 Shape Change of the Central Spindle in Anaphase—Anterior View in Polarized Light......Page 424 DISCUSSION......Page 426 Central Spindle Splaying......Page 435 REFERENCES......Page 436 The following note was added by Shinya Inoue in September of 2006:......Page 438 ABSTRACT......Page 440 MATERIALS AND METHODS......Page 441 Description of the Axostyle......Page 442 Movements of the Axostyle......Page 443 Wave Initiation other than at the Base......Page 449 Microbeam Experiments......Page 450 Cilia- and Flagella-like Movements of the Axostyle......Page 453 Perturbations of Axostyle Motility......Page 455 The Position of Force Production in the Sawtooth Waves......Page 456 REFERENCES......Page 457 Article 34 Video enhancement of polarization-based microscope images (in J Cell Biol, 1981)......Page 460 Comparison of Visual, Photographic, and Video Imaging......Page 461 Polarized Light Microscopy......Page 463 Differential Interference Contrast Microscopy......Page 466 DISCUSSION......Page 468 REFERENCES......Page 470 The Mitotic Spindle......Page 472 Models for Mitosis......Page 481 Coordination of Cytokinesis with Mitosis......Page 484 ACKNOWLEDGMENTS......Page 485 REFERENCES......Page 486 Article 36 Anoxia-induced gradient of cleavage inhibition (with Potrebic B et al, in J Cell Biol, 1982) with note added in 2006......Page 490 Article 37 Acrosomal reaction in Thyone sperm-I: Observation with high-resolution video microscopy (with Tilney LG, in J Cell Biol, 1982)......Page 498 Obtaining Sperm......Page 499 Interpretation of the Image......Page 502 The Periacrosomal Cup......Page 503 Interpretation of Structural Changes......Page 504 REFERENCES......Page 505 Article 38 Acrosomal reaction in Thyone sperm-II: Kinetics and mechanism (with Tilney LG, in J Cell Biol, 1982)......Page 506 Effect of Cytochalasin B on the Kinetics of Extension of the Acrosomal Process......Page 507 Changes in the Surface Morphology of the Acrosomal Process during Elongation......Page 508 Kinetics of Elongation of the Acrosomal Process is Consistent with the Diffusion of Actin to, and Their Assembly on Filaments at, the Tip of the Process......Page 511 Balance of Forces That Appear to Determine the Shape of the Growing Acrosomal Process......Page 512 REFERENCES......Page 513 Article 39 Acrosomal reaction in Thyone sperm-Ill: Actin assembly and water influx (with Tilney LG, in J Cell Biol, 1985)......Page 514 The Kinetics of Elongation of the......Page 515 Effect of Hypotonicity on the Kinetics of Extension of the Acrosomal Process......Page 516 The Effect of Salt on the Profilactin Region of Detergent-extracted Sperm......Page 520 The Assembly of Actin Is Sensitive to the Osmolarity of the Medium Which Surrounds the Sperm......Page 521 Preliminary Observations on the Ionic Composition of a Sperm Cell......Page 523 REFERENCES......Page 524 111.1. INTRODUCTION......Page 526 111.3. SOME BASIC FEATURES OF LIGHT WAVES......Page 527 111.4. OPTICAL ANISOTROPY......Page 529 111.5. BIREFRINGENCE......Page 530 111.6. DICHROISM......Page 533 111.7. DETERMINATION OF CRYSTALLINE AXES......Page 534 111.8. MOLECULAR STRUCTURE AND BIREFRINGENCE......Page 537 111.9. THE POLARIZING MICROSCOPE......Page 541 111.10. RECTIFICATION......Page 542 BIBLIOGRAPHY......Page 550 Article 41 Methods for microscopic observation of live gametes and embryos (with Lutz DA, in ''Methods in Cell Biology," 1986)......Page 560 II. Handling and Care of Animals......Page 561 III. Obtaining Gametes......Page 563 V. Fertilization and Culturing......Page 566 A. Removal of Fertilization Envelopes......Page 567 A. Microdrop Preparation......Page 569 B. Perfusion Chambers......Page 571 D. Manipulation Chambers......Page 572 A. Separation of Blastomeres......Page 573 B. Light Source......Page 574 C. Controlling Temperature during Observation......Page 575 E. Types and Selection of Optics......Page 576 F. Use of Video for Data Recording......Page 578 ACKNOWLEDGMENTS......Page 579 REFERENCES......Page 580 INTRODUCTION......Page 582 Optical Scrambler......Page 583 Image Processing......Page 584 Model Experiments......Page 585 Stereo Pair Generation from Serial Optical Sections......Page 586 Through-Focal Stereo Pairs......Page 587 Nonstereoscopic Through-Focal and Montaged Displays......Page 590 DISCUSSION......Page 591 REFERENCES......Page 593 Article 43 Asters in unequal cleavage of molluscs (with Dan K, in Intl J Repro and Develop, 1987)......Page 598 Introduction......Page 599 Materials and Methods......Page 600 A Working Hypothesis......Page 610 Polyspermic Eggs......Page 613 References......Page 615 Article 44 Automatic correction of spherical aberration in high NA microscope objectives (with Knudson RA and Inoue TD, in J Cell Biol, 1987)......Page 618 INTRODUCTION......Page 620 Micromanipulation......Page 621 Detachment of the Meiotic Apparatus From the Cell Cortex by Micromanipulation and Its Spontaneous Return to the Cortex......Page 622 Either Spindle Pole Can Establish an Association With the Attachment Site Cortex......Page 624 The MA Can Only Attach to the Cortex at One, Highly Localized Site......Page 626 DISCUSSION......Page 629 REFERENCES......Page 632 KD'S LAB......Page 634 JEAN'S HOMECOMING......Page 635 SPINDLE FIBERS......Page 637 MICROTUBULE DYNAMICS IN SPINDLE FIBERS......Page 638 AND NOW?......Page 641 REFERENCES......Page 642 I. Visualizing Objects Narrower than the Resolution Limit of the Light Microscope......Page 644 II. Diffraction Patterns of Very Narrow Objects......Page 648 III. Superresolution......Page 655 IV. Positional Information and Lateral Setting Accuracy......Page 660 V. Ultrathin Optical Sectioning and Axial Setting Accuracy......Page 663 VI. Other Factors Affecting the Choice of Pixel Dimensions, Calibration, and Choice of Magnification......Page 666 REFERENCES......Page 670 Article 48 Asymmetry of UV-microbeam severed microtubule ends (with Walker RA and Salmon ED, in J Cell Biol, 1989)......Page 672 Tubulin and Axoneme Preparation......Page 673 Results......Page 674 Discussion......Page 675 References......Page 677 Introduction......Page 680 Results......Page 681 Meiosis......Page 682 Discussion......Page 686 References......Page 691 The following note was added by Shinya Inoue in September of 2006:......Page 694 Article 51 Dynamics of mitosis and cleavage (in "Cytokinesis: Mechanism of Furrow Formation During Cell Division," 1990)......Page 696 ROLE OF MICROTUBULES IN SPINDLE POSITIONING......Page 698 OBSERVATIONS ON SPINDLE BEHAVIOR, POLAR BODY FORMATION, AND UNEQUAL CLEAVAGE IN SPISULA......Page 701 CLEAVAGE IN CYTOCHALASIN-TREATED ARBACIA EGGS......Page 704 ACKNOWLEDGMENTS......Page 706 REFERENCES......Page 708 Article 52 Microtubule breakdown in vivo visualized by high-speed video (with Febvre J et al, in Abstract of IX Intl Congr Protozool, 1993)......Page 710 Introduction......Page 712 Measurements......Page 713 Discussion......Page 718 References......Page 720 I. INTRODUCTION......Page 722 A. Resolution and Contrast......Page 723 B. Video Imaging......Page 724 C. Digital Image Processing......Page 725 III. IMAGE RESOLUTION AND CONTRAST IN CONVENTIONAL AND CONFOCAL LIGHT MICROSCOPY......Page 727 IV. ULTRATHIN OPTICAL SECTIONS WITH CONVENTIONAL LIGHT MICROSCOPY......Page 729 A. Practical Considerations......Page 730 B. Comments on Video and Image Processing Equipment......Page 737 V. EXAMPLES OF THREE- AND FOUR-DIMENSIONAL IMAGING AND ANALYSIS WITH CONVENTIONAL LIGHT MICROSCOPY......Page 739 ACKNOWLEDGMENTS......Page 742 REFERENCES......Page 743 Article 55 Recollection of Kayo Okazaki (in Develop Growth & Differ, 1994) with note added in 2006......Page 746 Article 56 A tribute to Katsuma Dan (in Biol Bull, 1994)......Page 750 Lateral Resolution*......Page 754 Axial Resolution......Page 756 CONFOCAL IMAGING......Page 757 Nipkow Disk......Page 758 Impact of Modern Video......Page 759 Laser Illumination......Page 760 Laser-Illuminated Confocal Microscopes......Page 761 CONFOCAL LASER-SCANNING MICROSCOPE......Page 762 Speed of Image or Data Acquisition......Page 763 Depth of Field in Phase-Dependent Imaging......Page 764 Lens Aberration......Page 765 Unintentional Beam Deviation......Page 766 SUMMARY......Page 767 REFERENCES......Page 768 1. EARLY HISTORY: THE DYNAMIC EQUILIBRIUM MODEL......Page 772 2. MICROTUBULE STRUCTURE, POLARITY, AND DYNAMIC INSTABILITY......Page 774 3. KINETOCHORE AND CHROMOSOME MOVEMENT ARE COUPLED TO MICROTUBULE ASSEMBLY/DISASSEMBLY......Page 778 4. KINETOCHORE MOTILITY AND RELATED FORCES MAY BE GENERATED BY MOTOR PROTEINS, BY MICROTUBULE ASSEMBLY/DISASSEMBLY, OR BY BOTH......Page 780 6. IN VITRO EVIDENCE FOR PULLING BY MICROTUBULE DISASSEMBLY......Page 782 7. THE NATURE OF MICROTUBULE TIP ATTACHMENT COMPLEXES......Page 783 8. MODELS FOR THE MICROTUBULE ASSEMBLY/DISASSEMBLY ENGINE......Page 784 9. THE KINETOCHORE AS A GOVERNOR......Page 785 10. CONCLUDING REMARKS......Page 787 REFERENCES......Page 788 8.1 Introduction......Page 794 8.2 Fabrication of test targets......Page 795 8.2.2 Phase target......Page 797 8.2.3 Fluorescent target......Page 799 8.2.4 Tilted test target......Page 800 8.3.1 Contrast transfer of coherent confocal vs. incoherent nonconfocal optics......Page 801 8.3.2 Contrast transfer of oil-immersion versus water-immersion objective......Page 803 8.3.3 Contrast transfer of bright-field microscope using tilted test targets......Page 805 8.3.4 Imaging square grids with the scanning force microscope......Page 807 References......Page 808 INTRODUCTION......Page 810 Cells and Cultures......Page 811 Imaging Technique......Page 812 Chemicals and Probes......Page 814 Induced Feet in Aggregating Amoebae......Page 815 Cytoskeletal Organization of the Feet......Page 817 Is Eupodium a True Foot?......Page 819 Cytoskeletal Organization of the Eupodium......Page 821 CONCLUSIONS......Page 823 REFERENCES......Page 824 Article 61 Photodynamic effect on Eosin-B-stained sperm (with Tran P and Burgos MH, in Biol Bull, 1997)......Page 826 Literature Cited......Page 827 Article 62 Video essay: Polarization microscopy of microtubule dynamics in mitotic spindle (with Oldenbourg R, in Mol Biol Cell, 1998)......Page 828 REFERENCES......Page 832 Article 63 Windows to dynamic fine structures, then and now (in FASEB J, 1999)......Page 836 REFERENCES......Page 840 Article 64 How well can amoeba climb? (with Fukui Y et al, in Proc Natl Acad Sci, USA, 2000)......Page 842 Article 65 EM of fertilization-induced changes in stratified Arbacia eggs (with Burgos M and Goda M, in Biol Bull, 2000)......Page 848 Literature Cited......Page 849 1.1. Early history: why centrifuge cells?......Page 850 1.2. Centrifuge microscopes......Page 851 1.3. Advantage of polarized light microscopy......Page 854 2.2. Mercury arc source......Page 855 2.4. Stroboscopic illumination......Page 856 2.5. Elimination oj laser speckle......Page 857 2.7. Design of the specimen chamber......Page 858 3.1. Image resolution......Page 861 3.4. Limiting factors......Page 862 References......Page 864 1. Stratification of Arbacia eggs......Page 866 3. Stratification and fragmentation of Chaetoptems oocytcs......Page 867 4. Ca2 ' activation of Chaetopterus oocytes......Page 868 5. Stratification of mouse oocytes (in collaboration with Drs Lin Liu and David Keefe of Marine Biological Laboratory, Woods Hole, and Women and Infants Hospital of Rhode Island)......Page 870 7. Stalling of directional amoeboid movement in myosin mutants of Dictyostelium (in collaboration with Dr Yoshio Fukui, Northwestern University Medical School)......Page 871 8. Haemoglobin crystals in toadtish-sickled red blood cells (in collaboration with Dr Ference I. Harosi, Marine Biological Laboratory, Woods Hole, and New College of the University of South Florida)......Page 872 9. Birefringence in fd virus solution (in collaboration with Drs Seth Fraden and Zvonimir Dogic, Brandeis University)......Page 873 Conclusion......Page 874 Acknowledgements......Page 875 References......Page 876 Literature Cited......Page 878 2 General Description of Polarization Aberrations of an Axial Beam Passing Through Lens Surfaces......Page 880 3.2 Uncoated Lenses......Page 883 4.1 Rotated Polarizer Technique......Page 884 4.2 Rotated Compensator Technique......Page 885 5 Rectification of Depolarization in a High-NA Microscope......Page 886 6 Experimental Results......Page 888 7 Conclusions......Page 890 Materials and Methods......Page 892 INTRODUCTION......Page 898 Sign of Birefringence and Coefficient of Birefringence......Page 899 Edge Birefringence......Page 900 Dichroism......Page 901 Colloidal Gold, Pinholes......Page 902 VIDEO MICROSCOPY......Page 903 Centrifuge Polarizing Microscope......Page 904 APPLICATIONS OF POLARIZATION MICROSCOPY......Page 905 LITERATURE CITED......Page 922 Article 72 Direct-view high-speed confocal scanner: CSU-10 (with Inoue TD, in "Methods in Cell Biology," 2002)......Page 926 I. Introduction......Page 927 II. Overview of Confocal Microscopes......Page 929 III. Design of the CSU-10......Page 930 IV. Sample Biological Applications......Page 932 V. Low Bleaching of Fluorescence Observed with the CSU-10......Page 940 VI. Very-High-Speed Full-Frame Confocal Imaging with the CSU-10......Page 943 VII. Haze Removal, Image Sharpening, and Dynamic Stereo Image Generation by Digital Signal Processing......Page 944 A. Neighborhood-Based Processing......Page 946 B. Sharpening and Unsharp Masking......Page 947 C. Arithmetic Precision......Page 950 D. Significance of Unsharp Masking......Page 951 F. From Image Stacks to Projections......Page 952 G. Viewing of 3-D and 4-D Images......Page 954 H. Storage Requirements......Page 955 VIII. Mechanical and Optical Performance of the CSU-10......Page 956 IX. Potentials of the CSU-10......Page 957 X. Addendum A (April, 2002)......Page 961 XI. Addendum B (personal communication from Dr. Kenneth R. Spring, June, 2002)......Page 963 References......Page 964 Article 73 Address by Emperor Akihito and acceptance speech by Shinya Inoue: International Prize for Biology (2003)......Page 970 1. Introduction......Page 976 2. Principles of Orientation-Independent DIC Microscopy......Page 977 3. Experimental Verification......Page 980 4. Conclusions......Page 984 References and Notes......Page 985 Article 75 Direct observation of red blood cells during centrifugation (with Hoffman JF, in Proc Natl Acad Sci USA, 2006)......Page 986 Shinya Scope-1......Page 992 Shinya Scope-2......Page 993 Shinya Scope-3......Page 994 Shinya Scope-4......Page 996 Shinya Scope-5 (and -6)......Page 998 Shinya Scope-6......Page 1000 Centrifuge Polarizing Microscope (CPM)......Page 1002 Appendix II: Curriculum Vita......Page 1006 Appendix III: List of Primary Publications......Page 1010 This book collects the publications of Shinya lnoué, pioneering cell biophysicist and winner of the 2003 International Prize for Biology. The articles cover the discovery, and elucidate the behavior in living cells, of the dynamic molecular filaments which organize the cell and play a central role in cell division. Other articles report on the development of microscopes, especially those using polarized light and digital image enhancement, which make possible studies of the ever-changing molecular architecture directly in living cells. This book also contains many high quality photo-micrographs as well as an appended DVD with an extensive collection of video movies of active living cells. After training in Tokyo and at Princeton University, Dr Inoué has held teaching positions at the University of Washington, Tokyo Metropolitan University, University of Rochester, Dartmouth Medical School, and University of Pennsylvania. He is a member of the U.S. National Academy of Sciences and currently holds the title of Distinguished Scientist at the Marine Biological Laboratory in Woods Hole, Massachusetts.