Development of Precise Aiming System of Laser Microbeam Cell Micromanipulation

激光微束细胞显微操作精密瞄准系统的研制

• 批准号: 62870072
• 负责人: YAMAMOTO Hajime
• 金额: $ 4.61万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62870072/
• 关键词: Laser; Micromanipulation; Bioengineerning; Cell biology

A novel biotechnique of cell micromanipulation by laser microbeam is pioneered. The system was essentially an inverted microscope with a thermostat interfaced to a He-Ne laser, a Ar' laser, a Nd:YAG laser and other kinds of laser, employed for microbeam cell irradiation, Each laser beam was guided by flexible optical fiber which could connect between several laser apparatus, microscope and laser beam selector. By a laser beam selector, the desired laser was selected to be irradiated to the specimen. The phase-contrasted image of the specimen was visually observed on a TV monitor screen and registered in a video tape recorder and a frame memory for a long time after the laser irradiation. The focal spot of the laser through a dichromatic mirror was superimposed on the specimen image and the focused irradiation at any desired site of the specimen was given by moving mechanical stage. To get a good aim at the intracellular target, such as one of organellae, the pahse-contrasted image of the cells was computerized by an image processor and was visually observed on a monitor screen. Laser microbeam micromanipulation was designed on a monitor screen by a interactive computer system with a mouse input. The laser beam was guided to the shtter controller which released the shutter of the laser for given period of time. The focused laser spot size of 10 mum in diameter was available by a 10-fold objective. The mechanical stge movement was accomplished by a prezonelectric motor which was good for a ultramicromovement.The present system of laser microbeam cell vaicromanipulation is probably more useful for studying the effects of laser irradiation on various type of cells than the in vivo systems. And our system has many advantages for detailed investigation of the effects on laser irradiation on the subcellular level of organella.

提出了一种利用激光微束进行细胞显微操作的新技术。该系统实质上是一个倒置的显微镜，带有恒温器，与He-Ne激光器、Ar’激光器、Nd：YAG激光器和其他类型的激光器接口，用于微束照射细胞。通过激光束选择器，选择所需的激光照射到样品。在TV监视器屏幕上目视观察样品的相位对比图像，并在激光照射后长时间记录在录像机和帧存储器中。激光通过分色镜的焦点叠加在样品图像上，并通过移动机械平台在样品的任何所需位置处给出聚焦照射。为了更好地瞄准细胞内的靶点，如器官的靶点，通过图像处理器将细胞的对比图像计算机化，并在监视器屏幕上进行视觉观察。激光微束微操作是通过鼠标输入的交互式计算机系统在显示器屏幕上设计的。激光束被引导到快门控制器，该快门控制器在给定的时间段内释放激光器的快门。10 μ m直径的聚焦激光光斑尺寸可通过10倍物镜获得。该系统的机械运动是由一个适合于超微运动的预驱动电动机完成的，它可能比在体系统更适合于研究激光照射对各种类型细胞的影响。该系统在研究激光对细胞器亚细胞水平的影响方面具有许多优势。

项目成果

茅野照雄: Transactiones Societatis Pathologicae Japonicae. 76. 122 (1987)

Teruo Kayano：日本病理学会汇刊。76. 122 (1987)

茅野照雄: 病態生理. 7. 229-234 (1988)

茅野辉夫：病理生理学。7. 229-234 (1988)

Suzuki,J.: Transactiones Societatis Pathologicae Japonicae. 77. 79 (1988)

铃木，J.：日本病理学会汇刊。

Kayano, Teruo: "Development of laser microbeam cell manipulation and its bioengineering application" Transactiones Societatis Pathologicae Japonicae. 76. 122 (1987)

Kayano Teruo：“激光微束细胞操作的发展及其生物工程应用”Transactiones Societatis Pathologicae Japonicae。

Suzuki, Junko: "Effects of argon ion laser on melanogenesis of human melanoma cell(HMV- II)" Transactiones Societatis Pathologicae Japomicae. 77. 79 (1988)

Suzuki, Junko：“氩离子激光对人黑色素瘤细胞（HMV-II）黑色素生成的影响”Transactiones Societatis Pathologicae Japomicae。