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Dual polarisation interferometry

 

Dual polarisation interferometry

Dual polarization interferometry (DPI) is an analytical technique that can probe molecular scale layers adsorbed to the surface of a waveguide by using the evanescent wave of a laser beam confined to the waveguide. It is typically used to measure the conformational change in proteins, or other biomolecules, as they function (referred to as the conformation activity relationship).

DPI[1] focuses laser light into two waveguides. One of these functions as the "sensing" waveguide having an exposed surface while the second one functions to maintain a reference beam. A two-dimensional interference pattern is formed in the far field by combining the light passing through the two waveguides. The DPI technique rotates the polarization of the laser, to alternately excite two polarization modes of the waveguides. Measurement of the interferogram for both polarizations allows both the refractive index and the thickness of the adsorbed layer to be calculated. The polarization can be switched rapidly, allowing real-time measurements of chemical reactions taking place on a chip surface in a flow-through system. These measurements can be used to infer conformational information about the molecular interactions taking place, as the molecule size (from the layer thickness) and the fold density (from the RI) change. DPI is typically used to characterise biochemical interactions by quantifying any conformational change at the same time as measuring reaction rates, affinities and thermodynamics.

The technique is quantitative and real-time (10 Hz) with a dimensional resolution of 0.01 nm.[2]

A novel application for Dual Polarization Interferometry emerged in 2008, where the intensity of light passing through the waveguide is extinguished in the presence of crystal growth. This has allowed the very earliest stages in protein crystal nucleation to be monitored.discussion page was opened in April 2012 for existing and potential users to network and provide information.

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Further reading

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