The long-term objectives of this project are to decipher the molecular and fine-structural organization of living cells which are responsible for mitotic chromosome movement and related cell motility. We propose to tackle this problem: through polarization optical and related methods of quantitative light microscopy by which we can follow the state of molecular assemblies and ordered fine structure as they dynamically appear and change, directly in living cells; and by developing the instruments and new methods which make feasible these investigations on living cells in real time, as the cells divide normally or under the influence of altered physical, physiological, or pharmacological conditions. Specifically, we will: (1) extend the application of video imaging and digital processing to improve the microscope resolution, image quality, speed, and capacity to enhance and display selected optical features in the live specimen, under conditions of known image reliability; (2) examine the mechanisms of mitotic and related cell motility primarily in the astral system that draws the female and male pronuclei together after fertilization anticipating their fusion, and in mitotic and meiotic spindles during induced microtubule depolymerization and reassembly; (3) advance the means for modulating selected reactions in targeted local sites within a living cell by the use of a minute, focused spot of light which activates or inactivates a photosensitive compound. Cell division, mitosis, and nuclear migration are fundamental, epochal events in the life of every cell. Without their regular progress, our tissues would lose their ability for normal growth, proper differentiation, or balanced sustenance. The technological contributions arising out of this project should also find timely applications in clinical as well as general and industrial microscopy.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Method to Extend Research in Time (MERIT) Award (R37)
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Cellular Biology and Physiology Subcommittee 1 (CBY)
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Marine Biological Laboratory
Woods Hole
United States
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Inoue, S (1997) The role of microtubule assembly dynamics in mitotic force generation and functional organization of living cells. J Struct Biol 118:87-93
Fukui, Y; Inoue, S (1997) Amoeboid movement anchored by eupodia, new actin-rich knobby feet in Dictyostelium. Cell Motil Cytoskeleton 36:339-54
Inoue, S (1996) Mitotic organization and force generation by assembly/disassembly of microtubules. Cell Struct Funct 21:375-9
Krendel, M; Inoue, S (1995) Anaphase spindle dynamics under D2O-enhanced microtubule polymerization. Biol Bull 189:204-5
Fukui, Y; Inoue, S (1995) Chemotaxis, aggregation behavior, and foot formation in Dictyostelium discoideum amoeba controlled by microbeam uncaging of cyclic-AMP. Biol Bull 189:198-9
Oldenbourg, R; Mei, G (1995) New polarized light microscope with precision universal compensator. J Microsc 180:140-7
Stemmer, A (1995) A hybrid scanning force and light microscope for surface imaging and three-dimensional optical sectioning in differential interference contrast. J Microsc 178:28-36
Inoue, S; Inoue, T D (1994) Through-focal and time-lapse stereoscopic imaging of dividing cells and developing embryos in DIC and polarization microscopy. Biol Bull 187:232-3
Roegiers, F; Tran, P; Inoue, S (1994) Mitosis, cleavage, and development of highly compressed sea urchin (Lytechinus variegatus) zygotes. Biol Bull 187:240-1
Oldenbourg, R; Terada, H; Tiberio, R et al. (1993) Image sharpness and contrast transfer in coherent confocal microscopy. J Microsc 172:31-9

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