研发了一套基于连续扫描的激光共焦扫描显微镜(laser confocal scanning microscopy,LCSM)系统。该系统采用工作台连续运动方式实现扫描,提出了利用单次采集的数据滤除随机噪声的方法,避免了多帧取平均对成像速度造成的影响。实现连续扫描的关键在于解决工作台运动与数据采集的同步问题,利用采集卡有限采集模式,合理匹配工作台参数和采集参数,成功解决了这一问题。详细介绍了影响分辨率的因素,通过合理选取探测器针孔直径,取样间隔,确保了实现高分辨率的要求。系统利用Visual C#开发的控制平台,成功地对生物细胞进行了扫描成像。实验结果表明:基于连续扫描的LCSM具有较高的分辨率,生成的显微图像没有任何畸变,并且成像速度有了大幅度提高。

A continuous scanning method and a new method of filtering random noise by single acquisition are brought forward to develop a laser confocal scanning microscopy (LCSM) system. The synchronous problem between scanning and data acquisition is solved by limited acquisition mode and rational cooperation of stage parameter and acquisition parameter. Factors of effect on resolution is also analyzed in detail. To meet the requirement of high resolution, the detector pinhole diameter and the sampling interval are properly picked up. The system takes images of the cells successfully by using the control platform based on Visual C#. The experiment results show that LCSM based on continuous scanning method provides a higher resolution, the generated microscopic image is absolutely undistorted.