Excitation and emission enhancement by using the plasmon mode formed on a plasmonic chip was studied with a microscope and micro-spectroscope. Surface plasmon resonance wavelengths were observed on one-dimensional (1D) and two-dimensional (2D) plasmonic chips by measuring reflection and transmission spectra, and they were assigned to the plasmon modes predicted by the theoretical resonance wavelengths. The excitation and emission enhancements were evaluated using the fluorescence intensity of yellow-green fluorescence particles. The 2D grating had plasmon modes of kgx45(2) (diagonal direction with m = 2) in addition to the fundamental mode of kgx(1) (direction of a square one side) in the visible range. In epifluorescence detection, the excitation enhancement factors of kgx(2) on the 1D and 2D chips were found to be 1.3-1.4, and the emission enhancement factor of kgx45(2) on the 2D chip was 1.5-1.8, although the emission enhancement was not found on the 1D chip. Moreover, enhancement factors for the other fluorophores were also studied. The emission enhancement factor of kgx(1) was shown to depend on the fluorescence quantum yield. The emission enhancement of 2D was 1.3-fold larger than that of 1D considering all azimuth components, and the 2D pattern was shown to be advantageous for bright fluorescence microscopic observation.
Keywords: fluorescence microscopy; microspectroscopy; plasmon enhancement; plasmon mode; surface plasmon resonance.