Development of Precise Aiming System of Laser Microbeam Cell Micromanipulation

提出了一种利用激光微束对细胞进行微操作的新型生物技术。该系统本质上是一个倒置显微镜，带有恒温器，连接He-Ne激光器、Ar激光器、Nd:YAG激光器等多种激光器，用于微束细胞照射，每束激光由柔性光纤引导，该光纤可连接多个激光设备、显微镜和激光束选择器。通过激光束选择器，选择所需的激光照射到样品上。在激光照射后，在电视屏幕上目视观察到试样的相对比图像，并在录像机和帧存储器中记录了很长时间。通过二色镜将激光的焦点光斑叠加在样品图像上，通过移动机械台对样品的任意位置进行聚焦照射。为了更好地瞄准细胞内的目标，如细胞器，细胞的相位对比图像由图像处理器计算机化，并在监视器屏幕上直观地观察。采用鼠标输入的交互式计算机系统，在显示器屏幕上设计了激光微光束微操作系统。激光束被引导到快门控制器，该控制器在给定的时间内释放激光器的快门。通过10倍物镜可获得直径为10微米的聚焦激光光斑。机械舞台的运动是由一个有利于超微运动的预区电机完成的。目前的激光微束细胞操纵系统可能比体内系统更适合于研究激光照射对各种类型细胞的影响。该系统为深入研究激光照射对细胞器亚细胞水平的影响具有许多优点。

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.