Systems and methods for fluorescence detection with a movable detection module

• US 20040224317A1
• Filed: 05/08/2003
• Published: 11/11/2004
• Est. Priority Date: 05/08/2003
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: MJ Research Incorporated (Bio-Rad Laboratories Incorporated)
• Inventors: Finney, Michael J., Kordunsky, Igor, Goldman, Jeffrey A.
• Granted Patent: US 7,148,043 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0829 Multi-well plates; Microtitration plates
• CPC Class Codes: B01L 2300/1822 using Peltier elements
• 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 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 21/276 with alternation of sample and standard in optical path
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2201/0446 Multicell plate, sequential

Abstract

A fluorescence detection apparatus for analyzing samples located in a plurality of wells in a thermal cycler and methods of use are provided. In one embodiment, the apparatus includes a support structure attachable to the thermal cycler and a detection module movably mountable on the support structure. The detection module includes one or more channels, each having an excitation light generator and an emission light detector both disposed within the detection module. When the support structure is attached to the thermal cycler and the detection module is mounted on the support structure, the detection module is movable so as to be positioned in optical communication with different ones of the plurality of wells. The detection module is removable from the support structure to allow easy replacement.

Claims

1. A fluorescence detection apparatus for analyzing samples located in a plurality of wells in a thermal cycler, the apparatus comprising:

a support structure attachable to the thermal cycler; and

a detection module movably mountable on the support structure, the detection module including;

an excitation light generator disposed within the detection module; and

an emission light detector disposed within the detection module;

wherein, when the support structure is attached to the thermal cycler and the detection module is mounted on the support structure, the detection module is movable so as to be positioned in optical communication with different ones of the plurality of wells.

2. The fluorescence detection apparatus of claim 1, wherein the detection module includes a plurality of excitation light generators and a plurality of emission light detectors arranged to form a plurality of excitation/detection pairs.

3. The fluorescence detection apparatus of claim 2, wherein each emission light detector is configured to detect a different range of wavelengths.

4. The fluorescence detection apparatus of claim 2, wherein each excitation light generator is configured to generate light at a different range of wavelengths.

5. The fluorescence detection apparatus of claim 2, wherein the plurality of excitation/detection pairs are arranged such that each excitation/detection pair is simultaneously positionable in optical contact with a different one of the plurality of wells.

6. The fluorescence detection apparatus of claim 2, wherein the plurality of excitation/detection pairs are arranged such that when a first one of the excitation/detection pairs is positioned in optical contact with any one of the plurality of wells, a different one of the excitation/detection pairs is not in optical contact with any one of the plurality of wells.

7. The fluorescence detection apparatus of claim 1, wherein the detection module is detachably mounted on the support structure, thereby enabling a user to replace the detection module with a different detection module.

8. The fluorescence detection apparatus of claim 1, further comprising:

a calibration element disposed such that the detection module is movable so as to be positioned in optical communication with the calibration element,

wherein the calibration element provides a known fluorescence response.

9. The fluorescence detection apparatus of claim 8, wherein the calibration element is located between two or more of the plurality of wells.

10. The fluorescence detection apparatus of claim 1, wherein positioning of the detection module with respect to the wells is controlled by an external computer.

11. The fluorescence detection apparatus of claim 8, wherein the calibration element is located in an area peripheral to the plurality of wells.

12. The fluorescence detection apparatus of claim 1, wherein operation of the excitation light generator and the emission light detector is controlled by an external computer.

13. A method for detecting the presence of a target molecule in a solution, the method comprising:

preparing a plurality of samples, each containing a fluorescent probe adapted to bind to a target molecule;

placing each sample in a respective one of a plurality of sample wells of a thermal cycler instrument, the thermal cycler instrument having a detection module movably mounted therein, the detection module including an excitation/detection channel, the excitation/detection channel including an excitation light generator disposed within the detection module and an emission light detector disposed within the detection module;

stimulating a reaction using the thermal cycler instrument; and

scanning the plurality of sample wells to detect a fluorescent response by moving the detection module and activating the excitation/detection channel,

wherein during the step of scanning, the detection module is moved such that the excitation/detection channel is sequentially positioned in optical communication with each of the plurality of sample wells.

14. The method of claim 13 wherein the target molecule is a nucleic acid sequence.

15. The method of claim 14 wherein the reaction is a polymerase chain reaction (PCR).

16. The method of claim 13 wherein the step of scanning includes:

positioning the detection module such that the excitation/detection channel is in optical communication with one of the sample wells;

briefly activating the excitation light generator of the excitation/detection channel;

detecting a fluorescent response using the emission light detector of the excitation/detection channel; and

repositioning the detection module such that the excitation/detection channel is in optical communication with a different one of the sample wells.

17. The method of claim 16 wherein the detection module includes at least two excitation/detection channels.

18. The method of claim 17 wherein, when the detection module is positioned such that a first one of its excitation/detection channels is in optical communication with one of the sample wells, a second one of its excitation/detection channels is in optical communication with a different one of the sample wells.

19. The method of claim 17 wherein, when the detection module is positioned such that a first one of its excitation/detection channels is in optical communication with one of the sample wells, a second one of its excitation/detection channels is not in optical communication with any of the sample wells.

20. The method of claim 16 wherein during the step of scanning, the detection module is substantially continuously in motion.