Use of cyanine dyes with evanescent wave fiber optic biosensors

Linda A. Tempelman; Joel P. Golden; George P. Anderson; Frances S. Ligler

doi:10.1117/12.190964

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Abstract

Cyanine fluorescent dyes which can be coupled to proteins have recently become available. They are excited in the 500-700 nm range and have large extinction coefficients which make them good candidates for sensitive immunosensor applications. We evaluated three of these dyes (Cy3, Cy5, and Cy5.5) in a direct fluoroimmunoassay on evanescent wave fiber optic biosensors. The biosensors differed in laser excitation sources and emission filters in order to accommodate dye requirements. The assay consisted of rabbit anti-goat IgG immobilized fiber probes being exposed to fluorophorelabeled goat IgG (gIgG). Upon binding of the dye-conjugated protein to the fiber, a signal was generated. When using the 514 nm laser device, Cy3-gIgG gave a substantially larger signal than TRITC-gIgG. On the 650 nm laser device, Cy5-gIgG provided very good signal, while that for Cy5.5 was moderate. Cy5.5 produced an excellent response on the 670 nm laser device. Use of these dyes provides a mechanism for improving the fiber optic biosensor by changing the excitation/emission region to an area of low background for clinical and environmental samples.

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Linda A. Tempelman, Joel P. Golden, George P. Anderson, and Frances S. Ligler "Use of cyanine dyes with evanescent wave fiber optic biosensors", Proc. SPIE 2293, Chemical, Biochemical, and Environmental Fiber Sensors VI, (21 October 1994); https://doi.org/10.1117/12.190964

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