Method and System for Multiplex Genetic Analysis

Abstract

The present disclosure provides apparatus, systems and method for detecting separately and substantially simultaneously light emissions from a plurality of localized light-emitting analytes. A system according to exemplary embodiments of the present disclosure comprises a sample holder having structures formed thereon for spatially separating and constraining a plurality of light-emitting analytes each having a single nucleic acid molecule or a single nucleic acid polymerizing enzyme, a light source configured to illuminate the sample holder, an optical assembly configured to collect and detect separately and substantially simultaneously light emissions associated with the plurality of light emitting analytes. The system may further include a computer system configured to analyze the light emissions to determine the structures or properties of a target nucleic acid molecule associated with each analyte.

1 .- 60 . (canceled) 
     
     
         61 . A method of interrogating target molecules within optical confinement regions disposed upon a substrate, comprising:
 providing photo-activated coupling groups on the substrate, including within the optical confinement regions;   directing activating radiation at the substrate to selectively activate the photo-activated coupling groups within the optical confinement regions;   depositing target molecules over the surface of the substrate such that at least a portion of the target molecules are retained within the optical confinement regions by coupling with the activated photo-activated coupling groups;   removing uncoupled target molecules from the substrate that are not retained within the optical confinement regions; and   directing light toward the substrate at an angle off-axis with respect to normal of the substrate such that at least a portion of the light is propagated by total internal reflection through at least a portion of the substrate illuminating coupled target molecules within a multiplicity of optical confinement regions.