Molecular Interferometric Imaging for Biosensor Applications

Molecular interferometric imaging (MI2) is a common-path interferometric imaging technique for detecting protein binding to surfaces. The experimental metrology limit is 10 pm/pixel longitudinal resolution at 0.4-mum diffraction-limited lateral resolution, corresponding to 1.7 attogram of protein, which is only 8 antibody molecules per pixel, near to single-molecule detection. The scaling mass sensitivity at the metrology limit is 5 fg/mm. We demonstrate a protein microarray application in a 128-multiplex immunoassay. Assay applications include prostate specific antigen (PSA) at a detection limit of 60 pg/mL and the cytokine interleukin-5 (IL-5) at a detection limit of 50 pg/mL. Realtime binding assays using MI2 enable the study of reaction kinetics of antibodies exposed to antigen, and the binding of antibody Fc regions to protein G.