Abstract
We present a new method for multi-color fluoroimmunoassays based on directional surface plasmon-coupled emission (SPCE). SPCE is coupling of excited fluorophores with a nearby thin metal film (silver) resulting in strongly directional emission into the underlying glass substrate. The angle at which the radiation propagates through the prism depends on emission wavelength and makes possible measurement of multiple analytes using multiple emission wavelengths. We demonstrated this possibility using two antibodies labeled with different fluorophores, binding to an antigen protein immobilized on the silver surface. We observed independent emission at a different angle on the glass prism, resulting of the surface binding of each antibody. This methodology can be readily extended to 3 or more fluorophores. This technology presents opportunity to develop highly sensitive multiplex assay format for biological agents' detection.
Original language | English |
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Article number | 56 |
Pages (from-to) | 316-325 |
Number of pages | 10 |
Journal | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
Volume | 5692 |
DOIs | |
State | Published - 21 Jul 2005 |
Event | Advanced Biomedical and Clinical Diagnostic Systems III - San Jose, CA, United States Duration: 23 Jan 2005 → 26 Jan 2005 |
Keywords
- Directional fluorescence
- Fluorescence immunoassay
- Multi-color assay
- Surface plasmon-coupled emission