Highly sensitive protein detection using polarization of fluorescence and background light of multi-layered substrate

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Polarization of fluorescence and background light from a multi-layered substrate were firstly investigated in case that a polarized light was used as an excitation light. As a result, the fluorescence was found not to be polarized although the background light was polarized. Irradiation of TE excitation light and detection of fluorescence polarized perpendicular to the excitation light were employed for a newly developed fluorescence detection instrument. Approximately 100-fold sensitivity improvement for fluorescence detection was achieved with this instrument. Moreover, Cy3 labeled streptavidin could be detected with 24-fold sensitivity improvement using an instrument equipped with the polarized excitation light.