Experimental and theoretical evaluation of surface plasmon-coupled emission for sensitive fluorescence detection

• Published in: Journal of Biomedical Optics 2008-09, Vol.13 (5), p.054021-054028
• Main Authors: Trnavsky, Michal, Enderlein, Joerg, Ruckstuhl, Thomas, McDonagh, Colette, MacCraith, Brian D
• Format: Article
• Language: English
• Subjects: Algorithms; Computer Simulation; Coupling (molecular); Emission; Emission analysis; Fluorescence; fluorescence intensity; Fluorescent Dyes - analysis; Models, Chemical; Optimization; Plasmons; Platforms; Reproducibility of Results; Selectivity; Sensitivity and Specificity; Silver - analysis; Spectrometry, Fluorescence - methods; supercritical angle fluorescence (SAF); Surface Plasmon Resonance - methods; surface plasmon-coupled emission (SPCE)
• ISSN: 1083-3668; 1560-2281
• DOI: 10.1117/1.2978067

Surface plasmon-coupled emission (SPCE) is a phenomenon whereby the light emitted from a fluorescent molecule can couple into the surface plasmon of an adjacent metal layer, resulting in highly directional emission in the region of the surface plasmon resonance (SPR) angle. In addition to high directionality of emission, SPCE has the added advantage of surface selectivity in that the coupling diminishes with increasing distance from the surface. This effect can be exploited in bioassays whereby a fluorescing background from the sample can be suppressed. We have investigated, both theoretically and experimentally, the SPCE effect for a Cy5-spacer-Ag layer system. Both the angular dependence of emission and the dependence of SPCE emission intensity on Cy5-metal separation were investigated. It is demonstrated that SPCE leads to lower total fluorescence signal than that obtained in the absence of a metal layer. This is the first experimental verification of the reduction in SPCE intensity compared to the metal-free case. Our results are in a good agreement with theoretical models. The validation of the theoretical model provides a basis for optimizing biosensor platform performance, particularly in the context of the advantages offered by SPCE of highly directional emission and surface selectivity.