Development of Precise Aiming System of Laser Microbeam Cell Micromanipulation

开创了一种利用激光微束进行细胞微操作的新生物技术。该系统实质上是一个倒置显微镜，其恒温器与He-Ne激光器、Ar‘激光器、NdYAG光等激光器接口，用于微束细胞的照射，每束激光由柔性光纤引导，可连接多个激光装置、显微镜和激光选束器。通过激光光束选择器，选择所需的激光照射到样品上。在激光照射后，在电视监视器屏幕上目视观察样品的相衬图像，并将其记录在录像机和帧存储器中很长时间。激光通过双色镜的焦斑叠加在试件图像上，通过移动机械台对试件的任意位置进行聚焦照射。为了更好地瞄准细胞内的目标，如细胞器中的一个，细胞的部分对比图像被图像处理器计算机化，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监视器屏幕上设计的。激光束被引导到粉碎控制器，该控制器在给定的时间段内释放激光器的快门。通过10倍的物镜可以获得直径为10微米的聚焦激光光斑尺寸。目前的激光微束细胞显微操纵系统可能比活体系统更适用于研究激光照射对不同类型细胞的影响。并且我们的系统在详细研究激光照射对细胞器亚细胞水平的影响方面具有许多优点。

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.