Millimeter-scale slide optical waveguides (OWGs) show the potential to break the barrier of easy-to-use and versatility for total internal reflection (TIR) fluorescence technology. In this paper, multi-frequency structured illumination (SI) patterns resulting from the evanescent field (EF) on the surface of a millimeter-scale polymer slide OWG are observed by measuring the fluorescence intensity distribution of fluorescent dyes deposited on the top of the OWG. The frequency, intensity, and stability of the SI patterns show a strong dependence on the coupling angle of the incident light (changing with the incident position). The distribution of multi-frequency SI patterns in the frequency space is demonstrated for different numerical aperture (NA) imaging systems (NA = 0.3, 0.6, and 0.8), indicating the potential for enhanced resolution for low NA systems with a simple and cheap polymer slide.
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