Observation of Double-Stained African Green Monkey Kidney COS-7 Cells Using Total Internal Reflection Double-Channel Fluorescence Microscopy
LIU Xiao-chen1, GUAN Li-zhao2, MA Wan-yun1*, ZHANG Hong-quan2
1. Key Laboratory for Atomic and Molecular Nanosciences of Ministry of Education, Department of Physics, Tsinghua University, Beijing 100084, China 2. Laboratory of Molecular Cell Biology and Tumor Biology, Department of Anatomy & Histology and Embryology, School of Basic Medical Sciences, Peking University, Beijing 100191, China
Abstract:Using a Dual-View wavelength splitter, double-stained African green monkey kidney COS-7 cells, transfected with pEGFP-Myosin 15a and costained with Rhodamine-filopodia were observed based on an ICCD(intensified charge couple device) fluorescence micro-imaging systems. Total internal reflection fluorescence microscopy was used to observe the overexpression of Myosin 15a to the tips of the elongation filopodia. An approach to collecting fluorescence in two channels and avoiding spectra crosstalk was employed to observe Myosin 15a and filopodia distribution in African green monkey kidney COS-7 cells. High sensitivity TIRF technology and two channels imaging method provided a wide application in bio-medical studies.
[1] Zhang Hongquan, Berg J S, Li Zhilun, et al. Nature Cell Biology, 2004, 6(6): 523. [2] Sousa A D, Cheney R E. Trends in Cell Biology, 2005, 15(10): 533. [3] Axelrod D. Traffic, 2001, 2(11): 764. [4] Wang Y M, Austin R H, Cox E C. Physical Review Letters, 2006, 97(4): 048302. [5] Bowser D N, Khakh B S. Proceedings of the National Academy of Sciences, 2007, 104(1): 4212. [6] Dunne P D, Fernandes R A, McColl J, et al. Biophysical Journal, 2009, 97: L05. [7] LIN Dan-ying, LIU Xiao-chen, MA Wan-yun(林丹樱,刘晓晨,马万云). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2009, 29(6): 1581. [8] Yamauchi K, Yang M, Jiang P, et al. Cancer Research, 2006, 66(8): 4208. [9] Ma B, He F, Jablonska J, et al. Microscopy Research and Technique, 2007, 70: 171. [10] Garini Yuval, Young T, McNamara G. Cytometry Part A, 2006, 69A: 735.