Multi-wavelength immunoassays using surface plasmon-coupled emission

Evgenia Matveeva, Joanna Malicka, Ignacy Gryczynski, Zygmunt Gryczynski, Joseph R. Lakowicz

Research output: Contribution to journalArticlepeer-review

70 Scopus citations


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.

Original languageEnglish
Pages (from-to)721-726
Number of pages6
JournalBiochemical and Biophysical Research Communications
Issue number3
StatePublished - 16 Jan 2004


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