Autofluorescence Imaging of Living Yeast Cells with Deep-Ultraviolet Surface Plasmon Resonance

Abstract

Autofluorescence in living cells on aluminum thin film was excited with deep-ultraviolet surface plasmon resonance (deep-UV SPR). Deep-UV SPR under aqueous medium was excited with Kretschmann configuration by using a sapphire prism. Deep-UV SPR is one of the promising techniques for high-sensitive autofluorescence imaging of living cells without staining. Label-free observation is significant for the structural analysis of living cells. We demonstrated the high-sensitive autofluorescence imaging of living yeast cells with deep-UV SPR. We applied a high refractive index prism, such as sapphire, which is suitable for the observation of specimens in aqueous medium, to excite deep-UV SPR. Although typical autofluorescence from living cells is buried in background noise, deep-UV SPR enhances the autofluorescence signal. The deep-UV SPR excitation of an aluminum thin film through a sapphire prism was investigated theoretically and experimentally. It showed that the fluorescence intensities are increased 2.8-fold. Deep-UV SPR enhanced the autofluorescence of cell structures, and yeast cells were found to be very sensitive. As a result, for water-immersed specimens, the sapphire-prism-based Kretschmann configuration excited SPR in deep-UV. Findings from this study suggest that deep-UV SPR can be considered an effective technique for attaining high-sensitivity observation of biological samples.