APPARATUS AND METHOD FOR DETECTION AND DISCRIMINATION MOLECULAR OBJECT

• US 20100256918A1
• Filed: 03/09/2010
• Published: 10/07/2010
• Est. Priority Date: 03/11/2009
• Status: Active Grant

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Li, Ming-Chia, Chen, Hung-Chun, Li, Yu-Tang, Chiou, Chung-Fan, Chu, Chang-Sheng
• Granted Patent: US 9,778,188 B2
• Time in Patent Office: Days
• US Class Current: 702/19
• CPC Class Codes: C12Q 1/6869 Methods for sequencing
• CPC Class Codes: C12Q 2537/157 A reaction step characterised by the number of molecules incorporated or released
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2565/607 being a sensor, e.g. electrode
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties
• CPC Class Codes: G01N 2015/1477 Multiparameters
• CPC Class Codes: G01N 2021/6441 with two or more labels
• 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 21/645 Specially adapted constructive features of fluorimeters

Abstract

An apparatus for detecting an object capable of emitting light. The apparatus comprises a light detector comprising at least two optical sensors capable of determining the intensity of the light; and a computer processing output signal generated by the optical sensors and comparing a result of the processing with a known result corresponding to a known type to determine whether the object belongs to the known type.

Claims

1. An apparatus for detecting an object capable of emitting light comprising:

(a) a light detector comprising at least a first optical sensor and a second optical sensor capable of determining the intensity of the light; and

(b) a computer processing the output signals generated by the optical sensors and comparing a result of the processing with a known result corresponding to a known type to determine whether the object belongs to the known type.

2. The apparatus of claim 1, further comprising:

(c) an optical structure collecting and directing a light emitted from the object to optical sensors.

3. The apparatus of claim 2, wherein the optical structure comprises a prism.

4. The apparatus of claim 2, wherein the optical structure comprises a grating.

5. The apparatus of claim 2, wherein the optical structure comprises a lens.

6. The apparatus of claim 2, wherein the optical structure comprises an optical filter.

7. The apparatus of claim 1, wherein the at least two optical sensors are stacked.

8. The apparatus of claim 1, wherein the at least two optical sensors are arranged horizontally.

9. The apparatus of claim 1, wherein the optical sensors are photodiodes.

10. The apparatus of claim 7, wherein the at least two optical sensors are formed by at least two stacking p-n junctions within one device.

11. The apparatus of claim 1, wherein the light detector comprises a circuit of signal amplifier, A/D convertor, integrator, comparator, logic circuit, readout circuit, memory, microprocessor, clock, and/or address.

12. The apparatus of claim 1, wherein processing the signals comprises:

calculating a sum of the signals;

comparing the sum with a threshold value to determine the presence of an object, wherein

when the sum is equal to or larger than the threshold value, it is determined that an object is present, and

when the sum is smaller than the threshold value, it is determined that an object is absent.

13. The apparatus of claim 1, wherein processing the signals comprises:

calculating a sum of the signals;

comparing the sum with a threshold value to determine the presence of an object, wherein

when the sum is smaller than the threshold value, it is determined that an object is present, and

when the sum is equal to or larger than the threshold value, it is determined that an object is absent.

14. The apparatus of claim 1, wherein

processing the signals comprises calculating a ratio of the signals,

comparing the results with the known result comprises comparing the calculated ratio with a known ratio corresponding to a known type, and

when the difference between the calculated ratio and the known ratio is within a certain range, it is determined that the object belongs to the known type.

15. The apparatus of claim 1, wherein the computer further compares the result of the processing with a plurality of known results corresponding to a plurality of known types to determine to which type the object belongs.

16. The apparatus of claim 12, wherein the computer further compares the calculated ratio with a plurality of known ratios corresponding to a plurality of known types to determine to which type the object belongs.

17. The apparatus of claim 14, wherein a known type having a known ratio that has the smallest difference to the calculated ratio among the plurality of known ratios is determined to be the type to which the object belongs.

18. The apparatus of claim 1, wherein the object comprises at least one quantum dot, at least one fluorescent protein, or at least one fluorescent small chemical moiety.

19. The apparatus of claim 1, wherein the object is fluorescently labeled and comprises at least one oligonucleotide, polynucleotide, oligopeptide, polypeptide, oligosaccharide, polysaccharide, or lipid.

20. The apparatus of claim 1, wherein the object is a single molecule.

21. The apparatus of claim 1, wherein the object comprises a target object without light emitting capability and a labeling object with light emitting capability.

22. The apparatus of claim 1, wherein the object comprises a target object without light emitting capability and a group of labeling objects with light emitting capability.

23. The apparatus of claim 1, wherein the object comprises only a single fluorophore chosen from a quantum dot, a fluorescent protein, and a fluorescent small chemical moiety.

24. The apparatus of claim 1, wherein the light detector is one of at least one light detector for detecting a plurality of objects, the plurality of objects are comprised by a liquid suspension, and the suspension flows past the at least one light detector.

25. A system for detecting a plurality of objects, the system comprising an array of the apparatuses of claim 1.

27. The method of claim 24, wherein the object comprises at least one quantum dot, at least one fluorescent protein, or at least one fluorescent small chemical moiety.

28. The method of claim 24, wherein the object is fluorescently labeled and comprises at least one oligonucleotide, polynucleotide, oligopeptide, polypeptide, oligosaccharide, polysaccharide, or lipid.

29. The method of claim 24, wherein the object is a single molecule.

30. The method of claim 24, wherein the object comprises a target object without light emitting capability and a labeling object with light emitting capability.

31. The method of claim 24, wherein the object comprises a target object without light emitting capability and a group of labeling objects with light emitting capability.

32. The method of claim 24, wherein the object comprises only a single fluorophore chosen from a quantum dot, a fluorescent protein, and a fluorescent small chemical moiety.

33. The method of claim 24, further comprising detecting a plurality of objects using at least one light detector, wherein the plurality of objects are comprised by a liquid suspension, and the method further comprises flowing the suspension past the at least one light detector.

26. A method for detecting an object capable of emitting light comprising:

(a) providing at least two output signals generated by a light detector in response to the light emitted from the object;

(b) processing the output signals; and

(c) comparing at least one result of the processing with at least one known result corresponding to at least one known type to determine whether the object belongs to the at least one known type.

34. A non-transitory computer-readable medium encoded with a computer program product, the computer program product, when executed by a computer, instructing the computer to perform the steps of:

(a) processing output signals generated by at least a first optical sensor and a second optical sensor, the signals being generated in response to absorbing light emitted from an object; and

(b) comparing a result of the processing with a known result corresponding to a known type to determine whether an object belongs to the known type.

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

(a) performing single molecule nucleic acid sequencing of at least one nucleic acid molecule, wherein the single molecule nucleic acid sequencing leads to emission of light correlated to the identity of at least one base comprised by the nucleic acid;

(b) detecting the light with at least one light detector comprising at least a first optical sensor and a second optical sensor;

(c) processing output signals from the at least two optical sensors; and

(d) comparing at least one result of the processing with at least one known result corresponding to at least one known type to determine an identity of at least one base comprised by the nucleic acid.