Fundamental Research on Laser Irradiation Effect on Cell Biology

激光照射对细胞生物学影响的基础研究

• 批准号: 02045006
• 负责人: FUJII Kan-ichi
• 金额: $ 3.97万
• 依托单位: Faculty OF ENGINEERING, IBARAKI UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for international Scientific Research
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02045006/
• 关键词: Antibody; Cell; ELISPOT method; ILISPOT method; Linker-arm; Optogalvanic; Raman; オプトガルバニック分光; 細胞生物学; 癌細胞; 波長依存性; ラマン分光; スト-クス光; 自己蛍光(Autofluorescence); 細胞エネルギ-準安定状態; ディジタルイメ-ジ法; レ-ザ-光照射

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

本国际合作项目所取得的研究成果概括为以下三项。(1)抗体的可视化定量测量的成功及其在蛋白质生产率决策中的应用。(2)通过拉曼散射法对溶剂中的小颗粒进行初步定性测量的成功，目的是分选癌细胞。(3)提出了激光照射癌细胞复苏的模型，并通过拉曼散射与激光光电流光谱的比较进行了实验验证，结果（1）是基于能够定量测定蛋白质的ILISPOT（免疫荧光连接免疫斑点分析）方法。与特定蛋白质结合的抗体直接与合适的染料结合。染料被荧光显微镜中包含的光源照射的光激发。染色抗体发出的荧光被摄像机拍摄下来， ...更多信息 ng从发光强度和发射斑点的截面定量测量蛋白质。这是由于天野博士最初的研究，是他博士学位的主要部分。毕业于美国伯明翰亚拉巴马大学。在进一步的研究中，我们希望使用一种新的含有F的连接臂以及He-Cd^+白光激光器作为荧光显微镜的光源。结果（2）是基于与细胞分选机同样的拉曼散射激光光谱法的结果。可以认为，当内脏器官具有不同的颜色时，存在用于激发单个内脏器官的最佳波长和来自其的特定发射光谱。因此，使用由三原色组成的He-Cd^+白色光激光器，预期可以同时分选三种细胞。作为此目的的准备，在丙酮和硝基苯中观察了拉曼光谱，从而完成了测量系统。细胞将在不久的将来在下一阶段进行检查。结果（3）是一个新的主题诞生在这个项目的目标的道路上。根据本项目提出的癌细胞的产生机制，癌细胞的能态高于健康细胞的能态，对应于原子中的亚稳态，该亚稳态被禁止通过光学跃迁返回基态。如果亚稳态可以通过激光照射激发到上能级而释放，然后通过光学跃迁释放到基态，则癌细胞可以恢复为健康细胞。为了验证这一假设，有必要检测由比亚稳态更高的能级跃迁到基态所发射的荧光。少

项目成果

K.FUJII: "Green-Yellow lasers in Cubr-AgBr-Ne systems operating at 40 KHz of repetition frequebcy" IEEE J.Quantum Electronics. 27. 2437-2481 (1991)

K.FUJII：“Cubr-AgBr-Ne 系统中的绿-黄激光器以 40 KHz 的重复频率运行”IEEE J.Quantum Electronics。

M.Takashima, S.Okada, H.Nishiyama, M.Matsuda and K.Fujii: "Full color printer with a He-Cd^+ white-light laser" Review of Scientific Instruments. 62. 1238-1245 (1991)

M.Takashima、S.Okada、H.Nishiyama、M.Matsuda 和 K.Fujii：“使用 He-Cd^ 白光激光的全彩色打印机”科学仪器评论。

G.Y. Yan,K-I.Fujii and A.L.Schawlow: "Relaxation-oscillator detection of optogalvanic spectra" Optics Letters. 15. 142-144 (1990)

G.Y.

K.FUJII: "Relaxation-oscillator detection of optogalvanic spectra" Optics Letters Review of Scientific Insrruments. 15. 142-144 (1990)

K.FUJII：“光电化学光谱的弛豫振荡器检测”光学快报科学仪器评论。

K.Oouchi, M.Suzuki and K.Fujii: "Green-Yellow lasers in Cubr-AgBr-Ne systems operating at 40 KHz of repetition frequency" IEEE Journal of Quantum Electronics. 27. 2437-2481 (1991)

K.Oouchi、M.Suzuki 和 K.Fujii：“Cubr-AgBr-Ne 系统中的绿-黄激光器以 40 KHz 的重复频率运行”IEEE 量子电子杂志。