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| home > ebr > spring 2003 > surface plasmon resonance applied to the high-throughput measurement of molecular binding |
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European BioPharmaceutical Review
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Basic Principles
Surface plasmon resonance (SPR) is an established optical technique for the detection of molecular binding, based on the generation of surface plasmons in a thin metal film (typically gold) that supports the binding chemistry. Surface plasmons are collective oscillations of free electrons constrained in the metal film.
These electrons are excited resonantly by a light field incident from a highly refractive index prism as shown in Figure 1. The angle of incidence q over which this resonant excitation occurs is relatively narrow, and is characterised by a reduction in the intensity of the reflected light which has a minimum at the resonant angle of incidence qr. The phase of the reflected light also varies nearly linearly with respect to q in this region.
The value of qr is a sensitive to the refractive index of the medium that resides within a few nanometres of the metal film. Small variations in the refractive index, due to the binding of a molecule to the film, may therefore be detected as a variation of this angle. In practice a thin layer of specific binding chemistry is deposited on the metal. Binding is then induced by flowing a fluid containing the target molecules over this composite surface.
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