激光扫描共聚焦显微镜及透射电镜观察肺癌A549细胞亚细胞结构溶酶体的对比分析  

Value of Laser Scanning Confocal Microscopy in Observing the Subcellular Structure-Lysosome of Lung Cancer A549 Cells:Compared with Transmission Electron Microscopy

作　　者：吴伟全[1] 王思捷[1] 李元歌[2] 何香红[1] 王永存[3] 吴平[1] 

机构地区：[1]广东医学院临床医学研究中心,广东湛江524001 [2]广东医学院附属医院放射科,广东湛江524001 [3]广东医学院附属医院肿瘤科,广东湛江524001

出　　处：《临床医学工程》2013年第6期666-668,共3页Clinical Medicine & Engineering

基　　金：广东省湛江市科技攻关项目(2012C3104028);广东医学院科研基金项目(M2011027)

摘　　要：目的比较分析激光扫描共聚焦显微镜和透射电子显微镜在观察肺癌A549细胞亚细胞结构溶酶体的运用价值。方法运用激光扫描共聚焦显微镜和透射电子显微镜分别观察肺癌A549细胞亚细胞结构溶酶体,实验分为三组:第一组A549细胞用透射电子显微镜进行观察、第二组A549细胞用激光扫描共聚焦显微镜获取细胞二维平面图像及进行细胞三维重构成像和第三组A549细胞用于激光扫描共聚焦显微镜的活细胞动态观察,并对结果进行比较分析。结果透射电子显微镜可以清晰、直观地观察到A549细胞亚细胞结构溶酶体的超微结构形态及数量;激光扫描共聚焦显微镜可以获取到分辨率高和成像清晰的二维平面溶酶体图像及三维重构溶酶体图像,可显示沿着Y轴旋转45°角的三维重构溶酶体成像,还可以动态观察A549细胞溶酶体荧光的变化情况,可相对定量分析溶酶体荧光强度。结论两种显微技术均可观察A549细胞亚细胞结构溶酶体的形态。与透射电子显微镜相比,激光扫描共聚焦显微镜更有利于反映溶酶体在A549细胞内的空间结构关系,并且有相对定量分析溶酶体荧光强度的优点。Objective To compare the value of laser scanning confocal microscopy and transmission electron microscopy in observing the subcellular structure-lysosome of lung cancer A549 cells. Methods The laser scanning confocal microscope and transmission electron microscope of the clinical medical research center were used to observe the lung cancer cell A549 subcellular structure-lysosomes, the experiment subjects were divided into three groups： the first group of A549 cells were observed by transmission electron microscope, second group of A549 cells were used by laser scanning confocal microscope to obtain cell 2D images and 3D reconstruction of cell imaging, and third groups of A549 cells were used for living cells dynamic observation by confocal laser scanning microscopy , then analyze the results. Results Transmission electron microscope could clearly, intuitively observe the ultrastructural morphology and quantity of A549 cells＇ subcellular structure-lysosomal; laser scanning confocal microscope could obtain the high resolution and clear 2D image and 3D reconstruction images of lysosomes, display the three-dimensional reconstruction of lysosome image along the Y axis rotated by an angle of 45°, dynamically observe the lysosomal in A549 cells, and relatively quantitative analyze the lysosomal fluorescence intensity. Conclusions The both techniques can be used to observe the subcellular structures lysosomal morphology in A549 cells. Compared with transmission electron microscopy, laser scanning confocal microscope have advantages in reflecting the spatial structure lysosomcs in A549 cells, and relative quantitative analyzing lysosomal fluorescence intensity.

关 键 词：激光扫描共聚焦显微镜 透射电子显微镜 A549 溶酶体 

分 类 号：R734.2[医药卫生—肿瘤]