He Ruei-Yu, Lin Chun-Yu, Su Yuan-Deng, Chiu Kuo-Chih, Chang Nan-Shan, Wu Hua-Lin, Chen Shean-Jen
Institute of Biomedical Engineering, National Cheng Kung University, Tainan 701, Taiwan.
Opt Express. 2010 Feb 15;18(4):3649-59. doi: 10.1364/OE.18.003649.
This paper demonstrates the first combination for wide-field surface plasmon (SP) phase microscopy and SP-enhanced fluorescence microscopy to image living cells' contacts on the surface of a bio-substrate simultaneously. The phase microscopy with a phase-shift interferometry and common-path optical setup can provide high-sensitivity phase information in long-term stability. Simultaneously, the fluorescence microscopy with the enhancement of a local electromagnetic field can supply bright fluorescent images. The combined microscope imposes a high numerical aperture objective upon the excitation of surface plasmon through a silver film with a thickness of 30 nm. The developed SP microscope is successfully applied to the real-time bright observation of the transfected fluorescence of living cells localized near the cell membrane on the bio-substrate and the high-sensitivity phase image of the cell-substrate contacts at the same time.
本文展示了首次将宽场表面等离子体(SP)相显微镜和SP增强荧光显微镜相结合,以同时对生物基质表面活细胞的接触进行成像。采用相移干涉测量和共光路光学装置的相显微镜能够在长期稳定性方面提供高灵敏度的相位信息。同时,通过增强局部电磁场的荧光显微镜可以提供明亮的荧光图像。组合显微镜在通过厚度为30nm的银膜激发表面等离子体时采用了高数值孔径物镜。所开发的SP显微镜成功地同时应用于对生物基质上靠近细胞膜定位的活细胞转染荧光的实时明亮观察以及细胞 - 基质接触的高灵敏度相位成像。