Label-free optical sensors

• US 10,365,224 B2
• Filed: 12/08/2008
• Issued: 07/30/2019
• Est. Priority Date: 12/06/2007
• Status: Active Grant

• Current Assignee: Genalyte, Inc.
• Original Assignee: Genalyte, Inc.
• Inventors: Gunn, III, Lawrence Cary
• Primary Examiner(s): Forman, Betty J
• Application Number:
  US12/746,747
• Publication Number:
  US 20110045472A1
• Time in Patent Office: 3,886 Days
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: G01N 21/7746 the waveguide coupled to a cavity resonator
• CPC Class Codes: G01N 21/84 Systems specially adapted for particular applications
• CPC Class Codes: G01N 33/54373 involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
• CPC Class Codes: G01N 33/574 for cancer

Abstract

Techniques, apparatus and systems are described for performing label-free monitoring of processes. In one aspect, a label-free monitoring system includes an array of label-free optical sensors to detect an optical signal in response to synthesis of one or more target genetic structures. Each label-free optical sensor is functionalized with a respective target genetic structure. The system also includes a fluid flow control module that includes fluid receiving units to provide paths for different fluids to flow into the fluid flow control module and at least one switch connected to the fluid receiving units to selectively switch among the fluid receiving units to receive a select sequence of the fluids through the fluid receiving units. The select sequence of the fluids includes at least a dNTP or base. A fluid channel is connected between the fluid flow control module and the array of sensors to allow the select sequence of the fluids to flow from the fluid flow control module to the array of label-free optical sensors.

Claims

1. A system for determining the nucleotide sequence of a target nucleic acid molecule, comprising:

a substrate;

a lower cladding layer provided over, and being supported by, said substrate;

one or more coupling waveguides disposed over said lower cladding layer to input an optical signal;

an array of label-free optical sensors provided over said lower cladding layer, each of said label-free optical sensors including a waveguide formed into a ring to serve as a ring resonant cavity, each of said ring resonant cavities in proximity to at least one of said one or more coupling waveguides with a spacing that provides an evanescent coupling region between said coupling waveguide and said ring resonant cavity, each label-free optical sensor being configured to modulate an optical signal in response to a nucleotide extension reaction, wherein each label-free optical sensor includes a sensing region that overlaps said evanescent coupling region and is functionalized with a target nucleic acid molecule, and wherein at least a portion of the nucleotide sequence of the target nucleic acid molecule is unknown, and wherein said label-free optical sensors of the array are each configured to produce an optical evanescent field in the sensing region such that when a single nucleotide is added to the target nucleic acid molecule via the nucleotide extension reaction, the label-free optical sensor modulates the optical signal to thereby provide a detectable indication of addition of the single nucleotide to the target nucleic acid molecule;

a fluid flow control module comprising fluid receiving units to provide paths for different fluids to flow into the fluid flow control module;

at least one switch connected to the fluid receiving units to select a fluid for flowing through the fluid receiving units, wherein the fluid comprises at least a nucleotide base; and

a fluid channel connected between the fluid flow control module and the array of label-free optical sensors to allow the selected fluid to flow from the fluid flow control module to the array of label-free optical sensors.

2. The system of claim 1, wherein the resonant cavity has a resonant frequency that shifts in response to the nucleotide extension reaction.

3. The system of claim 1, wherein the array of label-free optical sensors is configured to be interrogated.

4. The system of claim 1, wherein the resonant cavity has a complex refractive index of the resonant cavity that changes in response to the nucleotide extension reaction.

5. The system of claim 1, further comprising the fluid with a nucleotide base, the fluid having a single type of nucleotide base selected from the group consisting of A, G, C, T, and U.

6. The system of claim 1, further comprising a fluid which comprises a polymerase.

7. The system of claim 1, wherein the array of label-free optical sensors is configured to be interrogated while undergoing the nucleotide extension reaction.

8. The system of claim 1, wherein said substrate comprises a silicon-on-insulator wafer.

9. The system of claim 1, wherein the lower cladding layer comprises a buried insulator layer made of silicon dioxide.

10. The system of claim 1, wherein the at least one coupling waveguide comprises two coupling waveguides in evanescent coupling with said waveguide comprising said ring resonator cavity.

11. A system for determining the nucleotide sequence of a target nucleic acid molecule, comprising:

a substrate;

a lower cladding layer provided over, and being supported by, said substrate;

one or more coupling waveguides disposed over said lower cladding layer to input an optical signal;

a sensor array provided over said lower cladding layer, the sensor array comprising a plurality of optical resonators, each of said plurality of optical resonators being a waveguide formed in a closed waveguide loop, each of said optical resonators in proximity to at least one of said one or more coupling waveguides with a spacing that provides an evanescent coupling region between said coupling waveguide and said optical resonator;

an upper cladding layer provided over said plurality of optical resonators and said one or more coupling waveguides, said upper cladding layer including one or more cavities each at least partially surrounding a respective evanescent coupling region to serve as a sensing region, wherein each sensing region is functionalized with at least one target nucleic acid molecule, and wherein at least a portion of the nucleotide sequence of the target nucleic acid molecule is unknown, and wherein each resonator is configured such that, when a single nucleotide is added to the target nucleic acid molecule via a nucleotide extension reaction, the resonator detectably modulates the optical signal to thereby provide a detectable indication of addition of the single nucleotide to the target nucleic acid molecule;

a fluid flow control module; and

a flow channel in fluid communication with the sensor array, wherein the flow channel is configured to allow fluid to flow from the fluid flow control module to the sensor array.

12. The system of claim 11, wherein each resonant cavity has a resonant frequency that shifts in response to the nucleotide extension reaction.

13. The system of claim 11, wherein the sensor array is configured to be interrogated.

14. The system of claim 11, wherein each resonant cavity has a complex refractive index that changes in response to the nucleotide extension reaction.

15. The system of claim 11, wherein said closed waveguide loop comprises a closed waveguide loop of circular shape.

16. The system of claim 11, wherein said closed waveguide loop comprises a closed waveguide loop of non-circular shape.

17. The system of claim 11, wherein said closed waveguide loop has an irregular shape.

18. The system of claim 11, wherein said closed waveguide loop has an elliptical shape.