Substrates and optical systems and methods of use thereof having a single optically resolvable immobilized reaction component disposed within a nanometer-scale aperture
First Claim
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1. A method of detecting a signal from an analyte, comprising:
- a) providing a waveguide substrate comprising;
i) two or more optical waveguides disposed upon or within the waveguide substrate; and
ii) an analyte region disposed sufficiently proximal to a core of one of the two or more optical waveguides to be illuminated by an evanescent field emanating from the core when optical energy is passed through the core, wherein the analyte region comprises a single optically resolvable immobilized reaction component, and further wherein the analyte region is disposed within a nanometer-scale aperture that penetrates into a first surface on a first side of the waveguide substrate and extends toward the core,wherein the first surface is fluidically linked to the analyte region;
b) exposing the first surface to an analyte comprising a fluorescent or fluorogenic moiety that emits a signal in response to the evanescent field;
c) directing optical energy through the core, thereby generating the evanescent field and further illuminating the single optically resolvable immobilized reaction component; and
d) detecting a signal from the analyte upon interaction of the analyte with the single optically resolvable immobilized reaction component.
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Abstract
This invention provides substrates for use in various applications, including single-molecule analytical reactions. Methods for propagating optical energy within a substrate are provided. Devices comprising waveguide substrates and dielectric omnidirectional reflectors are provided. Waveguide substrates with improved uniformity of optical energy intensity across one or more waveguides and enhanced waveguide illumination efficiency within an analytic detection region of the arrays are provided.
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17 Claims
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1. A method of detecting a signal from an analyte, comprising:
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a) providing a waveguide substrate comprising; i) two or more optical waveguides disposed upon or within the waveguide substrate; and ii) an analyte region disposed sufficiently proximal to a core of one of the two or more optical waveguides to be illuminated by an evanescent field emanating from the core when optical energy is passed through the core, wherein the analyte region comprises a single optically resolvable immobilized reaction component, and further wherein the analyte region is disposed within a nanometer-scale aperture that penetrates into a first surface on a first side of the waveguide substrate and extends toward the core, wherein the first surface is fluidically linked to the analyte region; b) exposing the first surface to an analyte comprising a fluorescent or fluorogenic moiety that emits a signal in response to the evanescent field; c) directing optical energy through the core, thereby generating the evanescent field and further illuminating the single optically resolvable immobilized reaction component; and d) detecting a signal from the analyte upon interaction of the analyte with the single optically resolvable immobilized reaction component. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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Specification