全内角反射技术与转盘式共聚焦技术在细胞膜表面的成像比较

Comparison of TIRF and SDC on the Imaging of Cell Surface

转盘式共聚焦成像是一种高速、高分辨率成像技术,可以在高时间分辨率和空间分辨率的水平观察固定细胞内目标蛋白的分布及活细胞内目标蛋白的动态变化。全内角反射成像是一种观察距离玻片表面某个限定区域内蛋白质的分布和变化的成像技术,常用于观察固定细胞以及活细胞表面的亚细胞结构。该文以中性粒细胞和神经胶质瘤细胞作为观察对象,通过观测固定细胞膜表面蛋白质的分布以及追踪膜标记活细胞的动态变化对两种成像方法进行了比较。结果发现,就目前技术水平而言,二者均可以采集到清晰的细胞边缘,但全内角反射可以拍摄到更清晰的细胞膜表面结构,它在动态拍摄过程中光漂白相对较低,在快速捕捉过程中能够更加全面的捕捉到一个完整的运动过程。

Spinning disk confocal microscopy （SDC） is an imaging technique of high speed and resolution, and is a method to observe interested protein distribution in fixed cells and intracellular interested protein dynamics at high spatial and temporal resolution. A total internal reflection fluorescence （TIRF） microscope allows us to observe localization and dynamics of proteins in a restricted region from the interface of the coverslip and has been widely used for optical imaging of subcellular structure at the cell surface in both fixed and living cells. In this study, neutrophils and glioma cells were taken as observation objects, and two optical imaging approaches were compared on cell surface or live cell imaging. The of them can be used to get high resolution images of cell results indicate that, on the current technical level, both edges, but TIRF can capture higher-resolution images of cell surface and show lower photobleaching during Real-time imaging. Also, TIRF can help to observe an intact movement process better when capturing a rapid phenomenon.