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Total internal reflection fluorescence microscopy (TIRFM) induces the evanescent field from an incident light with an incident angle greater than the critical angle selectively to excite fluorescent molecules on or near a surface. The TIRFM not only provides enhanced understanding of cellular function but also improves signal-to-noise ratio of detecting signal in real time. However, fluorescent emission need to be increased when a dynamic biomolecular image is requested at the frame rate of greater than 100 frames/s. Therefore, the fluorescent signal is enhanced via surface plasmons to match the requirements of better efficiency and larger quantity. In this study, a plasmon-enhanced TIRFM whose operation is based on the electromagnetic field enhancement via surface and particle plasmon effects offered by a nano-scalar silver thin film and particles has been presented. The developed microscopy has been successfully used in the real-time observation of the enhanced fluorescence from the thrombomodulin protein of a living cell membrane. The simulated and
experimental results demonstrate that the plasmon-enhanced TIRFM can provide brighter living cell images through surface plasmon enhanced fluorescence.
R.-Y. He,K.-C. Chiu,Y.-D. Su,K.-L. Chang, andS.-J. Chen
"Study of cell adhesion and migration by using a plasmon-enhanced total internal reflection fluorescence microscope", Proc. SPIE 6088, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IV, 608802 (16 February 2006); https://doi.org/10.1117/12.647444
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R.-Y. He, K.-C. Chiu, Y.-D. Su, K.-L. Chang, S.-J. Chen, "Study of cell adhesion and migration by using a plasmon-enhanced total internal reflection fluorescence microscope," Proc. SPIE 6088, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IV, 608802 (16 February 2006); https://doi.org/10.1117/12.647444