Apparatus for single-molecule detection

• US 8,865,077 B2
• Filed: 06/11/2010
• Issued: 10/21/2014
• Est. Priority Date: 06/11/2010
• Status: Active Grant

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Chiou, Chung-Fan, Tsai, Rung-Ywan, Li, Yu-Tang, Shih, Chih-Tsung, Li, Ming-Chia, Chu, Chang-Sheng, Chung, Shuang-Chao, Chen, Jung-Po, Pu, Ying-Chih
• Primary Examiner(s): Warden, Jill
• Assistant Examiner(s): Kingan, Timothy G
• Application Number:
  US12/801,503
• Publication Number:
  US 20110306039A1
• Time in Patent Office: 1,593 Days
• Field of Search: 422 8203- 8211
• US Class Current: 422/82.11
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• 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: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01J 1/0425 using optical fibers
• CPC Class Codes: G01J 1/58 using luminescence generated by light
• 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: G02B 6/34 utilising prism or grating G02B6/293 takes precedence
• CPC Class Codes: Y10S 977/954 Of radiant energy
• CPC Class Codes: Y10S 977/957 Of chemical property or presence
• CPC Class Codes: Y10S 977/958 Of biomolecule property
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/145555 Hetero-N

Abstract

An apparatus for detecting an object capable of emitting light. The apparatus comprises a light source and a waveguide. The waveguide comprises a core layer and a first cladding layer. At least one nanowell is formed in at least the first cladding layer. The apparatus further comprises a light detector. The light detector can detect a light emitted from a single molecule object contained in the at least one nanowell.

Claims

1. A method of sequencing a nucleic acid, comprising the steps of:

providing a detection apparatus comprising;

a waveguide comprising;

a core layer; and

a first cladding layer made of a dielectric material;

wherein

at least one nanowell is formed in at least the first cladding layer, and

an upper opening of the at least one nanowell is larger than a bottom of the at least one nanowell;

a first opaque protection layer arranged over the first cladding layer without extending into the at least one nanowell;

a second opaque protection layer arranged below the waveguide, the second opaque protection layer having a nanostructured metal pattern; and

at least one light detector;

providing at least one nucleic acid molecule;

locating the at least one nucleic acid molecule individually within the at least one nanowell;

performing single molecule sequencing-by-synthesis of the at least one nucleic acid molecule, wherein the single molecule nucleic acid sequencing-by-synthesis leads to emission of light correlated to the identity of at least one base in the nucleic acid;

detecting the light with the detector, resulting in an output signal; and

processing the output signal to determine an identity of at least one base comprised by the nucleic acid.

2. The method of claim 1, wherein the single molecule nucleic acid sequencing-by-synthesis leads to emission of light via chemiluminescence.

3. The method of claim 1, wherein the detection apparatus further comprises a light source and the single molecule nucleic acid sequencing-by-synthesis leads to emission of light via fluorescence.