Integrated device with external light source for probing detecting and analyzing molecules

• US 10,288,566 B2
• Filed: 12/18/2017
• Issued: 05/14/2019
• Est. Priority Date: 08/08/2014
• Status: Active Grant

• Current Assignee: Quantum-Si Incorporated
• Original Assignee: Quantum-Si Incorporated
• Inventors: Rothberg, Jonathan M., Kabiri, Ali, Sickler, Jason W., Gyarfas, Brett J., Lackey, Jeremy, Schmid, Gerard, West, Lawrence C., Fife, Keith G., Cipriany, Benjamin, Ghasemi, Farshid
• Primary Examiner(s): Ayub, Hina F
• Application Number:
  US15/845,996
• Publication Number:
  US 20180120229A1
• Time in Patent Office: 512 Days
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 2021/6463 Optics
• CPC Class Codes: G01N 2021/6471 Special filters, filter wheel
• CPC Class Codes: G01N 21/6408 with measurement of decay time, time resolved fluorescence
• CPC Class Codes: G01N 21/6428 Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" in vivo A61B5/00; immunoassay G01N33/53
• CPC Class Codes: G01N 21/6454 using an integrated detector array
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 2201/064 Stray light conditioning
• CPC Class Codes: G01N 2201/0696 Pulsed
• CPC Class Codes: G01N 2201/08 Optical fibres; light guides
• CPC Class Codes: G01N 2201/12 Circuits of general importance; Signal processing

Abstract

System and methods for analyzing single molecules and performing nucleic acid sequencing. An integrated device includes multiple pixels with sample wells configured to receive a sample, which when excited, emits radiation. The integrated device includes at least one waveguide configured to propagate excitation energy to the sample wells from a region of the integrated device configured to couple with an excitation energy source. A pixel may also include at least one element for directing the emission energy towards a sensor within the pixel. The system also includes an instrument that interfaces with the integrated device. The instrument may include an excitation energy source for providing excitation energy to the integrated device by coupling to an excitation energy coupling region of the integrated device. One of multiple markers distinguishable by temporal parameters of the emission energy may label the sample and configuration of the sensor within a pixel may allow for detection of a temporal parameter associated with the marker labeling the sample.

Claims

1. An integrated device comprising:

a plurality of sample wells arranged on a surface of the integrated device;

at least one waveguide configured to deliver excitation energy to a vicinity of a portion of the plurality of sample wells;

a grating coupler positioned beneath the surface, wherein the grating coupler is configured to receive excitation energy incident on the surface from an external optical source and couple the excitation energy into the at least one waveguide; and

a reflector layer positioned to reflect light passing through the grating coupler towards the grating coupler,

wherein the reflector layer includes at least one opening, and the integrated device further comprises at least one monitoring sensor positioned to receive light through the at least one opening.

2. The integrated device of claim 1, wherein the grating coupler is positioned in a region of the integrated device separate from the plurality of sample wells.

3. The integrated device of claim 1, wherein the grating coupler has a width that tapers in a direction towards the at least one waveguide.

4. The integrated device of claim 1, wherein the at least one waveguide is tapered in a dimension perpendicular to a direction of light propagation along the at least one waveguide.

5. The integrated device of claim 1, wherein the integrated device further comprises:

a plurality of sensors configured to receive light from the plurality of sample wells.

6. The integrated device of claim 5, wherein the integrated device further comprises:

at least one baffle layer positioned between the plurality of sensors and the at least one waveguide, wherein the at least one baffle layer includes openings corresponding to individual sample wells of the plurality of sample wells.

7. The integrated device of claim 6, wherein the at least one baffle layer is further positioned between the grating coupler and a layer of the integrated device that includes the plurality of sensors.

8. The integrated device of claim 5, wherein the integrated device further comprises:

at least one filter positioned between the at least one waveguide and the plurality of sensors.

9. The integrated device of claim 8, wherein the at least one filter is configured to suppress transmission of the excitation energy.

10. The integrated device of claim 8, wherein the at least one filter includes a polarization filter.

11. The integrated device of claim 1, wherein the at least one monitoring sensor is configured to generate a signal indicative of alignment of an optical source to the grating coupler.

12. The integrated device of claim 1, wherein the integrated device further comprises at least one metal layer and the plurality of sample wells extend below the at least one metal layer.

13. The integrated device of claim 1, wherein individual sample wells of the plurality of sample wells have tapered sidewalls.

14. A method of forming an integrated device, the method comprising:

forming a plurality of sample wells arranged on a surface;

forming at least one waveguide configured to deliver excitation energy to a vicinity of a portion of the plurality of sample wells;

forming a grating coupler positioned beneath the surface, wherein the grating coupler is configured to receive excitation energy incident on the surface from an external optical source and couple the excitation energy into the at least one waveguide;

forming a reflector layer positioned to reflect light passing through the grating coupler towards the grating coupler, wherein the reflector layer includes at least one opening; and

forming at least one monitoring sensor positioned to receive light through the at least one opening.

15. The method of claim 14, wherein the method further comprises:

forming a plurality of sensors configured to receive light from the plurality of sample wells;

forming at least one baffle layer positioned between the plurality of sensors and the at least one waveguide; and

forming openings in the at least one baffle layer that correspond to individual sample wells of the plurality of sample wells.

16. The method of claim 14, wherein the method further comprises:

forming a plurality of sensors configured to receive light from the plurality of sample wells; and

forming at least one filter positioned between the at least one waveguide and the plurality of sensors.

17. The method of claim 14, wherein forming the plurality of sample wells further comprises forming at least one metal layer and forming the plurality of sample wells through the at least one metal layer to extend below the at least one metal layer.