激光扫描共聚焦显微镜的基本原理及其使用技巧  

The fundamentals and techniques in laser scanning confocal microscopy

作　　者：李叶[1,2,3] 黄华平[1,2] 林培群[1] 崔艳梅[1] 李勤奋[1,2] 郑勇奇[3] 

机构地区：[1]中国热带农业科学院环境与植物保护研究所,海南海口571101 [2]农业部热带农林有害生物入侵监测与控制重点开放实验室,海南儋州571737 [3]中国林业科学研究院林业研究所国家林业局森林培育重点试验室,北京100091

出　　处：《电子显微学报》2015年第2期169-176,共8页Journal of Chinese Electron Microscopy Society

基　　金：国家自然科学基金资助项目(No.31301629);中央级公益性科研院所基本科研业务费专项(No.2012hzs1J003);海南省重大专项(No.ZDZX2013023-1-25)

摘　　要：介绍了激光扫描共聚焦显微镜(laser scanning confocal microscopy,LSCM)的原理和构造,分析了LSCM的优点和缺点,并以花粉为例,详细介绍了共聚焦针孔直径、光电倍增管检测器增益、激光强度、扫描速度、扫描方式和Z轴步距等重要参数设置对共聚焦成像的不同影响。探讨了正确使用LSCM的方法与技巧,如获取高质量的图像、图像保存及图像处理,以便为科技人员利用LSCM开展更多植物学与环境科学相关的研究提供参考。The imaging principle of laser scanning confocal microscopy （ LSCM） was introduced in this paper. By taking the pollen as an example, the advantages and disadvantages of LSCM were discussed,and the effects of some important parameters including pinhole diameter,the gain of the photomultiplier tube detector, the intensity of the laser, scanning speed, scanning mode, Z-axis step on confocal imaging were introduced. In the meanwhile, the methods and techniques in LSCM well using were introduced, such as how to get high-quality images, images saving and image processing. These results will provide references for the researchers to carry out more botany and environmental science-related research by using LSCM.

关 键 词：激光扫描共聚焦显微镜 三维重建 花粉 针孔直径 激光强度 图像处理 

分 类 号：TP391.41[自动化与计算机技术—计算机应用技术]