The Study of Surface Plasmons Enhanced Total Internal Reflection Fluorescence Microscopy

• Main Authors: Hung-HsiuChen, 陳宏修
• Other Authors: Hsiang-Chen Chui
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/37755946975856426505

碩士 === 國立成功大學 === 光電科學與工程研究所 === 98 === In this thesis, we use the method of total internal reflection to excite surface plasmons on metal nanostructures, strengthening the regional electric field of metal nanoparticles to significantly enhance the fluorescent molecule signal. We also use nano-imprint process to fabricate various metal nanostructures as the large-area substrate to extensively enhance the fluorescent signal. Gold nanodisk and gold nanoparticle arrays are used for changing the sizes and interparticle spaces to study the optical enhancement properties of these structures, and the enhancement level of detected fluorescence signals. The traditional total internal reflection fluorescence microscopy (TIRFM) applies more than the critical angle of light incidence to generate evanescent wave (about several hundred nanometers) on the dielectric film where the fluorescent molecules are significantly stimulated. By this scheme, the TIRFM can achieve the resolution of nanometer level, which offers the image with very low fluorescence background noises and advancing the signal to noise ratio. However, in the real-time live cell imaging, the fluorescence signals from the cell membrane containing the dynamic information of specific molecular interactions are still needed to be enhanced. In order to get stronger fluorescence signals and to reduce the microscopic detection limit, we combine TIRFM and fabricated periodic metal nanostructure arrays to respectively measure fluorescent signals on the nano-imprint lithography-generated large-area polydimethylsiloxane (PDMS) substrates via localized surface plasmon (SP) excitations by means of the evanescent wave generation.