SYSTEMS AND METHODS FOR DETECTING MULTIPLE OPTICAL SIGNALS

• US 20100240063A1
• Filed: 06/01/2010
• Published: 09/23/2010
• Est. Priority Date: 03/10/2005
• Status: Active Grant

• Current Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Original Assignee: Gen-Probe Incorporated (Hologic Incorporated)
• Inventors: HAYES, Matthew J., Li, Haitao, KNIGHT, Byron J., Scalese, Robert F.
• Granted Patent: US 8,663,922 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6851 Quantitative amplification
• CPC Class Codes: C12Q 2537/165 Mathematical modelling, e.g. logarithm, ratio
• 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 2021/6432 Quenching
• 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 2035/00346 Heating or cooling arrangements
• CPC Class Codes: G01N 2035/00356 Holding samples at elevated temperature (incubation)
• CPC Class Codes: G01N 2035/00524 Mixing by agitating sample carrier
• CPC Class Codes: G01N 2035/0097 monitoring reactions as a function of time
• CPC Class Codes: G01N 2035/0441 for samples
• CPC Class Codes: G01N 2035/0443 for reagents
• CPC Class Codes: G01N 2035/0444 for cuvettes or reaction vessels
• CPC Class Codes: G01N 2035/0455 Coaxial carousels
• CPC Class Codes: G01N 2035/0491 Position sensing, encoding; closed-loop control
• CPC Class Codes: G01N 2035/103 using disposable tips
• 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 21/76 : Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 2201/0415 : Carrusel, sequential
• CPC Class Codes: G01N 35/00693 : Calibration
• CPC Class Codes: G01N 35/0092 : Scheduling
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: G01N 35/0099 : comprising robots or similar manipulators robots per se B25J
• CPC Class Codes: G01N 35/025 : having a carousel or turntable for reaction cells or cuvettes
• CPC Class Codes: G01N 35/026 : having blocks or racks of reaction cells or cuvettes
• CPC Class Codes: G01N 35/028 : having reaction cells in the form of microtitration plates
• CPC Class Codes: G01N 35/04 : Details of the conveyor system G01N35/021 - G01N35/028 take precedence
• CPC Class Codes: G01N 35/1002 : Reagent dispensers
• CPC Class Codes: Y10S 435/808 : Optical sensing apparatus
• CPC Class Codes: Y10S 435/809 : Incubators or racks or holders for culture plates or containers
• CPC Class Codes: Y10T 436/113332 : with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

To minimize cross talk in systems and methods for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, an excitation signal associated with each of the optical signals has a known excitation frequency, and any detected signal having a frequency that is inconsistent with the excitation frequency is discarded. The receptacles are moved relative to optical sensors configured to detect each unique optical signal from an associated receptacle, and to further minimize cross talk, the optical sensors are arranged so that only one reaction receptacle at a time is in a signal detecting position with respect to one of its associated optical sensors, and the optical sensors are grouped by the optical signal they are configured to detect so that a first optical signal is detected from each of the reaction receptacles before a second optical signal is detected from the reaction receptacles.

Claims

1. A method for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, said method comprising:

(a) directing an excitation signal at each reaction receptacle, the excitation signal having a known wavelength that excites an associated emission signal having a known emission wavelength, wherein the excitation signal has a known excitation frequency;

(b) detecting an emission signal emitted by each receptacle;

(c) detecting an emission frequency of the emission signal;

(d) discarding any detected emission signal having an emission frequency that is inconsistent with the known excitation frequency; and

(e) repeating steps (a)-(d) for each optical signal to be detected.

2. The method of claim 1, wherein the excitation frequency for one optical signal to be detected is different from the excitation frequency for at least one other optical signal to be detected.

3. The method of claim 2, wherein three or more different optical signals are detected from each of the reaction receptacles.

4. The method of claim 3, wherein the excitation frequencies are the same for two or more, but less than all, of the optical signals to be detected.

5. The method of claim 1, wherein the emission signal is a fluorescent emission.

6. The method of claim 1, wherein each different optical signal is generated by a different fluorescent dye contained in the reaction receptacle.

7. The method of claim 6, wherein the fluorescent dyes comprise rhodamine dyes in combination with a DABCYL quencher.

8. The method of claim 7, wherein the fluorescent dyes further comprise fluorescein dyes in combination with a DABCYL quencher.

9. The method of claim 8, wherein the dyes comprise two or more dyes selected from the group consisting of tetramethyl-6-rhodamine (“

TAMRA”

), tetrapropano-6-carboxyrhodamine (“

ROX”

), 6-carboxyfluorescein (“

FAM”

), and 2′

,7′

-dimethoxy-4′

,5′

-dichloro-6-carboxyrhodamin (“

JOE”

).

10. The method of claim 1,

wherein the two or more different optical signals comprise first, second, and third optical signals,

wherein the first optical signal comprises a first range of optical wavelengths, the second optical signal comprises a second range of optical wavelengths, and the third optical signal comprises a third range of optical wavelengths,

wherein the first and second range of optical wavelengths at least partially overlap each other, the second and third range of optical wavelengths at least partially overlap each other, but the first and third range of optical wavelengths do not overlap each other, and

wherein the excitation frequencies of the excitation signals associated with the first and third optical signals are the same and the excitation frequency of the excitation signal associated with the second optical signal is different from the excitation frequencies of the excitation signals associated with the first and third optical signals.

11. A system for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, wherein the reaction receptacles are disposed in a fixed arrangement with respect to each other, said system comprising:

(A) a plurality of optical sensors, each of said optical sensors being configured to detect one of the two or more optical signals and being associated with one of the reaction receptacles so that each reaction receptacle has associated therewith one optical sensor for each optical signal to be detected, whereby said optical sensors associated with each of the plurality of reaction receptacles are arranged in a configuration that enables each optical sensor to detect the associated optical signal emitted from the associated reaction receptacle;

(B) a transport mechanism constructed and arranged to effect relative movement between the plurality of reaction receptacles and the optical sensors to sequentially place each reaction receptacle into a signal-detecting position with respect to one of its associated optical sensors,

wherein

(1) said plurality of optical sensors are arranged so that only one reaction receptacle at a time can be in a signal-detecting position with respect to one of its associated optical sensors,

(2) optical sensors configured to detect each of the two or more optical signals are arranged in groups so that each of the reaction receptacles is placed in a signal-detecting position with respect to an associated optical sensor configured to detect one optical signal before any of the reaction receptacles is placed in signal-detecting position with respect to an associated optical sensor configured to detect a different optical signal,

(3) each optical sensor is configured to direct an excitation signal at an associated reaction receptacle that is in a signal-detecting position with respect to the optical sensor and to detect an emission signal emitted from the reaction receptacle that is excited by the excitation signal, and

(4) the optical sensors configured to detect a first emission signal are further configured to direct an excitation signal of a first excitation frequency at an associated receptacle and the optical sensors configured to detect a second emission signal are further configured to direct an excitation signal of a second excitation frequency at an associated receptacle, wherein the first and second excitation frequencies are different frequencies; and

(C) circuitry configured to discard any portion of the first emission signal having an emission frequency that is inconsistent with the first excitation frequency and to discard any portion of the second emission signal having an emission frequency that is inconsistent with the second excitation frequency.

12. The system of claim 11, wherein said plurality of optical sensors are disposed in fixed positions, and wherein said transport mechanism comprises a receptacle transport mechanism constructed and arranged to move the reaction receptacles into signal-detecting positions with respect to said fixed optical sensors.

13. The system of claim 12, wherein said receptacle transport mechanism comprises a rotatable carousel constructed and arranged for holding, moving, and positioning the reaction receptacles with respect to said optical sensors.

14. The system of claim 13, wherein said optical sensors are arranged at different angular positions with respect to said carousel.

15. The system of claim 13, wherein each of said reaction receptacles is positioned at a different radial position with respect to the axis of rotation of said carousel, and wherein the optical sensors associated with each reaction receptacle are positioned at different angular locations and at the radial position corresponding to the associated reaction receptacle.

16. The system of claim 15, further comprising baffles configured to optically isolate each of said optical sensors from the other optical sensors, wherein said baffles comprise walls arranged concentrically and at different radial positions with respect to the axis of rotation of said carousel so as to be located between radially-adjacent reaction receptacles.

17. The system of claim 15, wherein each of said reaction receptacles comprises a test tube, and wherein said test tubes are connected to each other in an aligned arrangement and an upright orientation and further wherein said connected arrangement of test tubes is carried on said carousel in a generally radial orientation so that each test tube is located at a different radial position.

18. The system of claim 14, wherein said system comprises three optical sensors associated with each reaction receptacle, each of the three optical sensors being configured to detect a different one of three different optical signals.

19. The system of claim 18, comprising fifteen optical sensors to detect three different optical signals emitted by each of at least five reaction receptacles.

20. The system of claim 19, wherein said optical sensors are equally spaced at 24 degree increments about said carousel.

21. The system of claim 11, comprising three optical sensors associated with each reaction receptacle, each of the three optical sensors being configured to detect a different one of three different optical signals.

22. The system of claim 11, wherein each of said reaction receptacles comprises a test tube, and wherein said test tubes are connected to each other in an aligned arrangement and an upright orientation.

23. The system of claim 11, further comprising baffles configured to optically isolate each of said optical sensors from the other optical sensors.

24. The system of claim 11, wherein each of said optical sensors comprises a fluorometer constructed and arranged to detect fluorescent emissions from the reaction receptacles.

25. The system of claim 11, wherein the two or more different optical signals are optical signals having different wavelengths.

26. A method for detecting two or more different optical signals emitted from each of a plurality of reaction receptacles, wherein the reaction receptacles are disposed in a fixed arrangement with respect to each other, said method comprising:

providing a plurality of optical sensors, each of said optical sensors being configured to detect one of the two or more optical signals and being associated with one of the reaction receptacles so that each reaction receptacle has associated therewith one optical sensor for each optical signal to be detected;

effecting relative movement between the plurality of reaction receptacles and the optical sensors to sequentially place each reaction receptacle into an signal-detecting position with respect to one of its associated optical sensors;

arranging the plurality of optical sensors so that only one reaction receptacle at a time can be in a signal-detecting position with respect to one of its associated optical sensors,

grouping the optical sensors configured to detect each of the two or more optical signals so that each of the reaction receptacles is placed in a signal-detecting position with respect to an associated optical sensor configured to detect one optical signal before any of the reaction receptacles is placed in signal-detecting position with respect to an associated optical sensor configured to detect a different optical signal.

directing an excitation signal at a unique excitation frequency with each optical sensor at an associated reaction receptacle that is in a signal-detecting position with respect to the optical sensor and detecting an emission signal emitted from the reaction receptacle that is excited by the excitation signal;

identifying an emission frequency of the emission signal detected from each excited reaction receptacle; and

discarding any portion of the emission frequency identified that is inconsistent with the excitation frequency of the excitation signal that was directed at the reaction receptacle .

27. The method of claim 26, wherein the plurality of optical sensors are disposed in fixed positions, and effecting relative movement comprises moving the reaction receptacles into signal-detecting positions with respect to the fixed optical sensors.

28. The method of claim 27, wherein moving the reaction receptacles comprises moving each of the reaction receptacles in a circular path.

29. The method of claim 28, wherein the optical sensors are arranged at different angular positions with respect to the circular path of the associated reaction receptacle.

30. The method of claim 28, wherein all of the reaction receptacles are moved in a circular path having a common axis of rotation and each of the reaction receptacles is positioned at a different radial position with respect to the axis of rotation, and wherein the optical sensors associated with each reaction receptacle are positioned at different angular locations and at the radial position corresponding to the associated reaction receptacle.

31. The method of claim 30, wherein each of the reaction receptacles comprises a test tube, and wherein the test tubes are connected to each other in an aligned arrangement and an upright orientation and further wherein moving the reaction receptacles in a circular path comprises carrying the connected arrangement of test tubes in a generally radial orientation with respect to the axis of rotation.

32. The method of claim 26, comprising providing three optical sensors associated with each reaction receptacle, each of the three optical sensors being configured to detect a different one of three different optical signals.

33. The method of claim 26, wherein each of the reaction receptacles comprises a test tube, and wherein the test tubes are connected to each other in an aligned arrangement and an upright orientation.

34. The method of claim 26, further comprising optically isolating each of the optical sensors from the other optical sensors.

35. The method of claim 26, wherein each of the optical sensors comprises a fluorometer constructed and arranged to detect fluorescent emissions from the reaction receptacles.

36. The method of claim 26, wherein each different optical signal is generated by a different fluorescent dye contained in the reaction receptacle.

37. The method of claim 36, wherein the fluorescent dyes comprise rhodamine dyes in combination with a DABCYL quencher.

38. The method of claim 37, wherein the fluorescent dyes further comprise fluorescein dyes in combination with a DABCYL quencher.

39. The method of claim 38, wherein the dyes comprise two or more dyes selected from the group consisting of tetramethyl-6-rhodamine (“

TAMRA”

), tetrapropano-6-carboxyrhodamine (“

ROX”

), 6-carboxyfluorescein (“

FAM”

), and 2′

,7′

-dimethoxy-4′

,5′

-dichloro-6-carboxyrhodamin (“

JOE”

).

40. The method of claim 26, wherein the two or more different optical signals are optical signals having different wavelengths.