Fundamental Research on Laser Irradiation Effect on Cell Biology

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The research results obtained by this International joint project are summarized in three items as follows.(1) Success of Visualized quantitative measurements of antibody and its application to decision of production rate of protein.(2) Success of preliminary qualitative measurements of small particles in a solvent by Ramman scattering method with an intent of sorting cancer cells.(3) A model of revival of cancer cell by laser irradiation was proposed and its experimental verification based on comparison of Ramman scat tering with optogalvanic laser spectroscopy is in progressing.The result (1) is based on the ILISPOT (Immunofluorescence Linked Immunospot Assay) method which enable the quantitative measurements of protein. The antibody bound to specific protein is directly combined with a suitable dye.the dye is excited by the light irradiated from the light-source contained in a fluorescence microscope. The fluorescence, emitted from the dyed-antibody id taken into a video camera, bei … More ng measured the protein quantitatively from the light emission intensity and the cross-section of emission spot. This is due to Dr.Amano's original investigation, being it major part of his Ph.D. thesis at The University of Alabama at Birmingham, U.S.A.. In further investigation, we would like to use a new linker-arm containing F as well as He-Cd^+ white-light laser as the light source of the fluorescence microscope. It is expected to improved the s/n ratio by this means.The result (2) is based on Raman scattering laser spectroscopy like in a cell sorter. It can be suggested that an optimal wavelength for exciting individual inner organ and a specific emission spectra from it would exist as the inner organs have different colors. So, using the He-Cd^+ white light laser consisting of three primary colors, it is expected that three kinds of cells can be sorted simultaneously. As a preparation for this aim, Ramman spectra were observed in acetone and nitrobenzine, thereby measuring system was accomplished. Cells would be examined in the near future at next stage.The result (3) is a new subject born on the way to the goal of this project. According to the Generation mechanism of the cancer cell proposed in this project, the energy state of a cancer cell is higher than that in a healthy cell, corresponding to a metastable state in an atom which is forbidden to return to the ground state Via optical transition. If the metastable state can be released via excitation to an upper level by a laser irradiation, then to the ground state via optical transition, the cancer cell can return to a healthy cell. For the verification to this hypothesis, it is necessary to detect the fluorescence emitted by the transition from higher level than metastable state onto the ground state. Less