Protein reactions with surface-bound molecular targets detected by oblique-incidence reflectivity difference microscopes.

We applied oblique-incidence reflectivity difference microscopes (a form of polarization-modulated nulling ellipsometry) to detection of biomolecular microarrays without external labeling in a study of protein reactions with surface-immobilized targets. We show that the optical reflectivity difference signals can be quantitatively related to changes in surface mass density of molecular layers as a result of the reactions. Our experimental results demonstrate the feasibility of using oblique-incidence reflectivity difference microscopes for high-throughput proteomics research such as screening unlabeled protein probes against libraries of surface-immobilized small molecules.