Automated Optical Reader for Nucleic Acid Assays

• US 6,597,450 B1
• Filed: 01/14/2000
• Issued: 07/22/2003
• Est. Priority Date: 09/15/1997
• Status: Expired due to Term

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Hansen, Timothy, OʼKeefe, Christian V., Failing, Frank L., Pope, Willard C., Andrews, Jeffrey P., Scrivens, Brian G.
• Primary Examiner(s): Smith, Zandra V.
• Application Number:
  US09/483,686
• Time in Patent Office: 1,285 Days
• Field of Search: 356/311, 356/437, 356/440, 356/246, 356/417, 356/317, 250/361.C, 422/52, 422/65
• US Class Current: 356/317
• CPC Class Codes: G01N 2021/6419 Excitation at two or more wavelengths
• CPC Class Codes: G01N 2021/6484 Optical fibres
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 35/028 having reaction cells in the form of microtitration plates

Abstract

An apparatus and method employing a plurality of light emitting devices which each can get light through a respective optical fiber toward a respective sample of a plurality of samples in a time-staggered manner. Light is generated in each of the samples at different times consistent with the times at which light is irradiated onto the sample. A single detector is used to detect the lights emitted from the plurality of samples at these different times. A plurality of bifurcated optical cable are coupled to the light emitting devices and single light detector, and the integrated end of each bifurcated cable acts as the light emitting port and light detecting port. Multiple targets can be detected from each of the plurality of samples in the same manner by providing an apparatus and method employing a different plurality of light emitting devices and single detector for each target to be detected.

Claims

1. An apparatus for analyzing a plurality of samples, comprising:

a first plurality of light emitting devices which are each controlled to radiate a first excitation light toward one sample of the plurality of samples at different first excitation times to excite a first probe contained in each of the plurality of samples to generate a first emission light at first emission times corresponding to said different first excitation times;

a first light detector which is controlled to detect said first emission light emitted from said first probe at said first emission times;

a second plurality of light emitting devices which are each controlled to radiate a second excitation light toward one sample of the plurality of samples at different second excitation times to excite a second probe contained in each of the plurality of samples to generate a second emission light at second emission times corresponding to said different second excitation times; and

a second light detector which is controlled to detect the second emission light emitted from said second probe at said second emission times.

2. An apparatus as claimed in claim 1, further comprising:

a first plurality of light emitting/detecting ports, each of which is;

(a) optically coupled to one of said first plurality of light emitting devices to emit said first excitation light emitted by said light emitting device toward one sample of the plurality of samples,

(b) configured to receive said first emission light from said first probe contained in said one of the plurality of samples, and

(c) optically coupled to said first light detector to radiate said first emission light emitted by said one of the plurality of samples to said first light detector; and

a second plurality of light emitting/detecting ports, each of which is;

(a) optically coupled to one of said second plurality of light emitting devices to emit said second excitation light emitted by said light emitting device toward one sample of the plurality of samples,

(b) configured to receive said second emission light from said second probe contained in said one of the plurality of samples, and

(c) optically coupled to said second light detector to radiate said second emission light emitted by said one of the plurality of samples to said second light detector.

3. An apparatus as claimed in claim 2, further comprising:

a first light sensing bar wherein said first plurality of light emitting/detecting ports are included on said first light sensing bar such that each one of said first plurality of light emitting/detecting ports is optically coupled to one of said first plurality of light emitting devices and optically coupled to said first light detector such that said first excitation light is radiated from said first plurality of light emitting devices toward the plurality of samples, and such that said first emission light is radiated from the plurality of samples towards said first light detector; and

a second light sensing bar wherein said second plurality of light emitting/detecting ports are included on said second light sensing bar such that each one of said second plurality of light emitting/detecting ports is optically coupled to one of said second plurality of light emitting devices and optically coupled to said second light detector such that said second excitation light is radiated from said second plurality of light emitting devices toward the plurality of samples, and such that said second emission light is radiated from the plurality of samples towards said second light detector.

4. An apparatus as claimed in claim 1, wherein said first excitation light and said second excitation light are excitation lights of different wavelengths.

5. An apparatus as claimed in claim 1, wherein said first excitation light has a wavelength within the blue light spectrum generating said first emission light having a wavelength within the green light spectrum and said second excitation light has a wavelength within the orange light spectrum generating said second emission light having a wavelength within the red light spectrum.

6. An apparatus as claimed in claim 2, wherein said plurality of light emitting/detecting ports comprises eight light emitting/detecting ports.

7. An apparatus as claimed in claim 1, wherein said first light detector is a photomultiplier tube and said second light detector is a photomultiplier tube.

8. An apparatus as claimed in claim 1, wherein said first probe is a detection probe for detecting one type of target nucleic acid and said second probe is a detection probe for detecting a different type of target nucleic acid.

9. An apparatus as claimed in claim 1, wherein said first probe is a detection probe for detecting target nucleic acid and said second probe is an internal amplification control probe.

10. An apparatus as claimed in claim 1, further comprising a controller which is adaptable to control:

said first light detector to detect the presence of light when said first plurality of light emitting devices are off, and which provides a first signal indicative of light detection when said first light detector detects the presence of light with said first plurality of light emitting devices off; and

said second light detector to detect the presence of light when said second plurality of light emitting devices are off, and which provides a second signal indicative of light detection when said second light detector detects the presence of light with said second plurality of light emitting devices off.

11. An apparatus as claimed in claim 2, wherein each of said first plurality of light emitting/detecting ports is optically coupled to one of said first plurality of light emitting devices and is optically coupled to said first light detector by a plurality of bifurcated light conducting devices and wherein each of said second plurality of light emitting/detecting ports is optically coupled to one of said second plurality of light emitting devices and is optically coupled to said second light detector by a plurality of bifurcated light conducting devices.

21. An apparatus as claimed in claim 1, further comprising:

a third plurality of light emitting devices which are each controlled to radiate a third excitation light toward one sample of the plurality of samples at different third excitation times to excite a third probe contained in each of the plurality of samples to generate a third emission light at third emission times corresponding to said different third excitation times; and

a third light detector which is controlled to detect said third emission light emitted from said third probe at said third emission times.

22. An apparatus as claimed in claim 21, wherein said third excitation light has a wavelength different from said first excitation light and different from said second excitation light.

23. An apparatus as claimed in claim 21, wherein said first, second and third probes are detection probes for detecting different types of target nucleic acid.

24. An apparatus as claimed in claim 21, wherein said first and second probes are detection probes for detecting different types of target nucleic acid and said third probe is an internal amplification control probe.

12. A method for analyzing a plurality of samples, comprising:

controlling each one of a first plurality of light emitting devices to radiate a first excitation light toward one sample of the plurality of samples at different first excitation times to excite a first probe contained in each of the plurality of samples to generate a first emission light at first emission times corresponding to said different first excitation times;

controlling a first light detector to detect said first emission light emitted from said first probe at said first emission times;

controlling each one of a second plurality of light emitting devices to radiate a second excitation light toward one sample of the plurality of samples at different second excitation times to excite a second probe contained in each of the plurality of samples to generate a second emission light at second emission times corresponding to said different second excitation times; and

controlling a second light detector to detect the second emission light emitted from said second probe at said second emission times.

13. A method as claimed in claim 12, further comprising:

controlling a first plurality of light emitting/detecting ports to emit said first excitation light, to receive said first emission light, and to radiate said first emission light to said first light detector;

wherein each one of said first plurality of light emitting/detecting ports is;

(a) optically coupled to one of said first plurality of light emitting devices to emit said first excitation light emitted by said light emitting device toward one sample of the plurality of samples,

(b) configured to receive said first emission light from said first probe contained in said one of the plurality of samples, and

(c) optically coupled to said first light detector to radiate said first emission light emitted by said one of the plurality of samples to said first light detector; and

controlling a second plurality of light emitting/detecting ports to emit said second excitation light, to receive said second emission light, and to radiate said second emission light to said second light detector;

wherein each one of said second plurality of light emitting/detecting ports is;

(a) optically coupled to one of said second plurality of light emitting devices to emit said second excitation light emitted by said light emitting device toward one sample of the plurality of samples,

(b) configured to receive said second emission light from said second probe contained in said one of the plurality of samples, and

(c) optically coupled to said second light detector to radiate said second emission light emitted by said one of the plurality of samples to said second light detector.

14. A method as claimed in claim 13, further comprising:

controlling a first light sensing bar to emit said first excitation light, to receive said first emission light, and to radiate said first emission light to said first light detector;

wherein said first plurality of light emitting/detecting ports are included on said first light sensing bar such that each one of said first plurality of light emitting/detecting ports is optically coupled to one of said first plurality of light emitting devices and optically coupled to said first light detector such that said first excitation light is radiated from said first plurality of light emitting devices toward the plurality of samples, and such that said first emission light is radiated from the plurality of samples towards said first light detector; and

controlling a second light sensing bar to emit said second excitation light, to receive said second emission light, and to radiate said second emission light to said second light detector;

wherein said second plurality of light emitting/detecting ports are included on said second light sensing bar such that each one of said second plurality of light emitting/detecting ports is optically coupled to one of said second plurality of light emitting devices and optically coupled to said second light detector such that said second excitation light is radiated from said second plurality of light emitting devices toward the plurality of samples, and such that said second emission light is radiated from the plurality of samples towards said second light detector.

15. A method as claimed in claim 12, further comprising:

controlling said first plurality of light emitting devices to radiate said first excitation light of a first wavelength; and

controlling said second plurality of light emitting devices to radiate said second excitation light of a second wavelength.

16. A method as claimed in claim 12, further comprising:

controlling said first plurality of light emitting devices to radiate said first excitation light of a blue wavelength and controlling said first light detector to receive said first emission light having a wavelength within the green light spectrum; and

controlling said second plurality of light emitting devices to radiate said second excitation light of a orange wavelength and controlling said second light detector to receive said second emission light having a wavelength within the red light spectrum.

17. A method as claimed in claim 12, further comprising:

controlling said first light detector to detect said first emission light radiated from said first probe; and

controlling said second light detector to detect said second emission light radiated from said second probe wherein said first and second probes are detection probes for detecting different types of target nucleic acid.

18. A method as claimed in claim 12, further comprising:

controlling said first light detector to detect said first emission light radiated from said first probe; and

controlling said second light detector to detect said second emission light radiated from said second probe wherein said first probe is a detection probe for detecting target nucleic acid and said second probe is an internal amplification control probe.

19. A method as claimed in claim 12, further comprising:

controlling a controller to control said first light detector to detect the presence of light when said first plurality of light emitting devices are off, and to provide a first signal indicative of light detection when said first light detectors detects the presence of light with said first plurality of light emitting devices off; and

controlling said controller to control said second light detector to detect the presence of light when said second plurality of light emitting devices are off, and to provide a second signal indicative of light detection when said second light detectors detects the presence of light with said second plurality of light emitting devices off.

20. A method as claimed in claim 19, further comprising:

adjusting the detecting sensitivity of the first light detector based on information provided by the first light detector indicative of the detected emission light when said first plurality of light emitting devices are off; and

adjusting the detecting sensitivity of the second light detector based on information provided by the second light detector indicative of the detected emission light when said second plurality of light emitting devices are off.

25. A method as claimed in claim 12, further comprising:

controlling each one of a third plurality of light emitting devices to radiate a third excitation light toward one sample of the plurality of samples at different third excitation times to excite a third probe contained in each of the plurality of samples to generate a third emission light at third emission times corresponding to said different third excitation times; and

controlling a third light detector to detect the third emission light emitted from said third probe at said third emission times.

26. A method as claimed in claim 25, further comprising:

controlling said first plurality of light emitting devices to radiate said first excitation light of a first wavelength;

controlling said second plurality of light emitting devices to radiate said second excitation light of a second wavelength; and

controlling said third plurality of light emitting devices to radiate said third excitation light of a third wavelength.

27. An apparatus for analyzing a plurality of samples, comprising:

at least three probes contained in each of the plurality of samples;

a plurality of light emitting devices and a light detector device for each of said at least three probes;

wherein each one of said at least three pluralities of light emitting devices is capable of radiating a different wavelength excitation light and wherein each of said at least three excitation lights is capable of exciting only one of said at least three probes to generate a corresponding emission light;

each of said light emitting devices in each of said at least three pluralities of light emitting devices are each controlled to radiate said at least three excitation lights toward one sample of the plurality of samples at different excitation times to excite one of said at least three probes to generate said at least three emission lights at emission times corresponding to said different excitation times; and

each of said at least three light detectors is controlled to detect said at least three emission lights emitted at said emission times.

28. A method for analyzing a plurality of samples wherein each of the plurality of samples contains at least three probes, comprising:

controlling a plurality of light emitting devices for each of said at least three probes to radiate a different wavelength excitation light toward one sample of the plurality of samples at different excitation times to excite only one of said at least three probes to generate an emission light for each of said at least three probes at emission times corresponding to said different excitation times; and

controlling a light detector for each of said at least three probes to detect said emission light emitted from each of said at least three probes at said emission times.