Matveeva Evgenia, Malicka Joanna, Gryczynski Ignacy, Gryczynski Zygmunt, Lakowicz Joseph R
Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201, USA.
Biochem Biophys Res Commun. 2004 Jan 16;313(3):721-6. doi: 10.1016/j.bbrc.2003.12.010.
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.
我们描述了一种使用表面等离子体耦合发射（SPCE）进行多波长免疫分析的新方法。这种现象是激发的荧光团与附近的薄金属膜（在我们的例子中是银）耦合，导致强烈的定向发射进入下面的玻璃基板。辐射通过棱镜传播的角度取决于相关波长的表面等离子体角度。这些角度取决于发射波长，从而允许使用多个发射波长测量多种分析物。我们使用用罗丹明红-X或AlexaFluor 647标记的抗体证明了这种可能性。这些抗体针对与银表面结合的抗原蛋白。每种标记抗体的发射在玻璃棱镜上以不同角度发生，从而允许独立测量每种抗体的表面结合。这种SPCE免疫分析方法可以很容易地扩展到4个或更多波长。
