Journal
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Volume 313, Issue 3, Pages 721-726Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2003.12.010
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Funding
- NCRR NIH HHS [RR-08119, P41 RR008119, P41 RR008119-14, P41 RR008119-13] Funding Source: Medline
- NIBIB NIH HHS [EB-00682, R01 EB000682-05, R21 EB000981, R01 EB000682, R01 EB000682-03, R21 EB000981-01, R01 EB000682-04, R21 EB000981-02, EB-00981] Funding Source: Medline
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We describe a new method for multi-wavelength immunoassays using surface plasmon-coupled emission (SPCE). This phenomenon is coupling of excited fluorophores with a nearby thin metal film, in our case silver, resulting in strongly directional emission into the underlying glass substrate. The angle at which the radiation propagate through the prism depends on the surface plasmon angle for the relevant wavelength. These angles depend on emission wavelength, allowing measurement of multiple analytes using multiple emission wavelengths. We demonstrated this possibility using antibodies labeled with either Rhodamine Red-X or AlexaFluor 647. These antibodies were directed against an antigen protein bound to the silver surface. The emission from each labeled antibody occurred at a different angle on the glass prism, allowing independent measurement of surface binding of each antibody. This method of SPCE immunoassays can be readily extended to 4 or more wavelengths. (C) 2003 Elsevier Inc. All rights reserved.
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