Apparatus and method for detection and discrimination molecular object

• US 9,778,188 B2
• Filed: 03/09/2010
• Issued: 10/03/2017
• Est. Priority Date: 03/11/2009
• Status: Active Grant

• Current Assignee: Industrial Technology Research Institute
• Original Assignee: Industrial Technology Research Institute
• Inventors: Chen, Hung-Chun, Li, Ming-Chia, Chu, Chang-Sheng, Li, Yu-Tang, Chiou, Chung-Fan
• Primary Examiner(s): Skibinsky, Anna
• Application Number:
  US12/720,352
• Publication Number:
  US 20100256918A1
• Time in Patent Office: 2,765 Days
• Field of Search: None
• 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. A system for detecting an object capable of emitting light comprising:

a nucleic acid object

a light detector for performing detection on the nucleic acid object, the light detector comprising a multi-junction photodiode which includes;

a first optical sensor including a first p-n junction and configured to generate a first output signal in response to the light; and

a second optical sensor including a second p-n junction and configured to generate a second output signal in response to the light,

wherein the first optical sensor is made of a first semiconductor material and the second optical sensor is made of a second semiconductor material different from the first semiconductor material, and the first and second optical sensors have different responsive properties; and

a computer configured to;

sum the first and second output signals to obtain a total signal,

divide the second output signal by the total signal to obtain a first ratio, compare the first ratio to a first range corresponding to a known type,

divide the first output signal by the second output signal to obtain a second ratio, and

confirm whether the object belongs to the known type by determining whether the second ratio is within a second range corresponding to the known type,

wherein the object is a nucleic acid.

2. The apparatus of claim 1, further comprising:

an optical structure collecting and directing a light emitted from the object to the 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 the object comprises at least one quantum dot, at least one fluorescent protein, or at least one fluorescent small chemical moiety.

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

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

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

16. 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.

17. 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.

18. 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.

27. The apparatus according to claim 1, further comprising a non-transitory computer-readable medium encoded with a computer program product, the computer program product, when executed by the computer, instructing the computer to perform the steps of:

receiving a first output signal generated by a first optical sensor in response to light emitted by an object and a second output signal generated by a second optical sensor in response to the light, the first and second optical sensors being made of different semiconductor materials and having different responsive properties;

summing the first and second output signals to obtain a total signal;

determining whether the total signal is equal to or larger than a threshold value;

if the total signal is equal to or larger than the threshold value, dividing the second output signal by the total signal to obtain a first ratio;

determining whether the first ratio is within a first range corresponding to a known type;

if the first ratio is within the first range, dividing the first output signal by the second output signal to obtain a second ratio; and

confirming whether the object belongs to the known type by determining whether the second ratio is within a second range corresponding to the known type.

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

a nucleic acid object

providing a light detector for performing detection on the nucleic acid object, the light detector comprising a multi-junction photodiode which includes;

a first optical sensor including a first p-n junction and configured to generate a first output signal in response to the light; and

a second optical sensor including a second p-n junction and configured to generate a second output signal in response to the light,

wherein the first optical sensor is made of a first semiconductor material and the second optical sensor is made of a second semiconductor material different from the first semiconductor material, and the first and second optical sensors have different responsive properties;

summing the first and second output signals to obtain a total signal;

dividing the second output signal by the total signal to obtain a first ratio;

compare the first ratio to a first range corresponding to a known type;

dividing the first output signal by the second output signal to obtain a second ratio; and

confirming whether the object belongs to the known type by determining whether the second ratio is within a second range corresponding to the known type,

wherein the object is a nucleic acid.

20. The method of claim 19, wherein the object comprises at least one quantum dot, at least one fluorescent protein, or at least one fluorescent small chemical moiety.

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

22. The method of claim 19, wherein the object is a single molecule.

23. The method of claim 19, wherein the object comprises a target object without light emitting capability and a labeling object with light emitting capability.

24. The method of claim 19, wherein the object comprises a target object without light emitting capability and a group of labeling objects with light emitting capability.

25. The method of claim 19, wherein the object comprises only a single fluorophore chosen from a quantum dot, a fluorescent protein, and a fluorescent small chemical moiety.

26. The method of claim 19, 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.