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Comparison of TIRF and SDC on the Imaging of Cell Surface
Yu Wenying1, Geng Guangfeng1, Zhang Chen2, Chen Ting1, Liang Haoyue1, Cheng Xuelian1, Bai Yang1, Yang Wanzhu1*
1State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China;
2Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin 300052, China
2Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin 300052, China
Abstract: 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 results indicate that， on the current technical level， both of them can be used to get high resolution images of cell 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.
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