In fluorescence microscopy, the upright microscope mounting a Xe lamp, fluorescent filter units including Cy5, GFP, and that composed of Cy3-excitation filter (λ = 535 ± 25 nm), Cy5-dichroic mirror (cutoff wavelength: 660 nm), Cy5-emission filter (λ = 700 ± 30 nm), and imaging cameras, was used. In cell imaging, two-color sensitive images of breast cancer cells were observed with a Bull’s eye-plasmonic chip with 480 nm-pitch. The fluorescence intensity in the Bull’s eye type was larger than those in the line & space pattern and hole array pattern under the microscope. On the plasmonic chip, EpCAM and EGFR in breast cancer cells were detected by 7-fold brighter fluorescence compared with those on the glass slide. In VSD imaging, action potential corresponding to spiking changes of membrane potential of neurons was observed with VSD. Rat hippocampal neurons were cultured in plasmonic-chips and glass-bottomed dishes and neurons were labeled with di-4 ANEPPDHQ. The frame rate for fluorescence images was 1ms, and the action potential was directly measured. Spikes of VSD signals were frequently detected on the plasmonic chip compared with glass-bottomed dish.

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Keiko Tawa, "Application of a plasmonic chip to sensitive bio-detection and fluorescence microscopic imaging (Conference Presentation)," Proc. SPIE 11082, Plasmonics: Design, Materials, Fabrication, Characterization, and Applications XVII, 110821C (Presented at SPIE Nanoscience + Engineering: August 15, 2019; Published: 9 September 2019); https://doi.org/10.1117/12.2526688.6083790557001.