SYSTEMS AND METHODS FOR FLUORESCENCE DETECTION WITH A MOVABLE DETECTION MODULE

• US 20070059754A1
• Filed: 11/01/2006
• Published: 03/15/2007
• Est. Priority Date: 05/08/2003
• Status: Active Grant

• Current Assignee: Bio-Rad Laboratories Incorporated
• Original Assignee: Bio-Rad Laboratories Incorporated
• Inventors: Finney, Michael, Kordunsky, Igor, Goldman, Jeffrey
• Granted Patent: US 7,749,736 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;

a shuttle movably mounted on the support structure; and

a detection module detachably attached to the shuttle, the detection module including;

a housing having an opening oriented toward the plurality of wells;

an excitation light generator disposed within the housing; and

an emission light detector disposed within the housing,

wherein, when the support structure is attached to the thermal cycler, the shuttle is movable to position the detection module in optical communication with different wells of the plurality of wells.

2. The fluorescence detection apparatus of claim 1 wherein an excitation optical path from the excitation light generator to the opening has a fixed length and a detection optical path from the opening to the emission light detector has a fixed length.

3. 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.

4. The fluorescence detection apparatus of claim 3 wherein the calibration element is located between two or more of the plurality of wells.

5. The fluorescence detection apparatus of claim 3 wherein the calibration element is located in an area peripheral to the plurality of wells.

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

7. 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.

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

a housing having an opening at one end thereof;

a fitting on an exterior surface of the housing, the fitting adapted to attach the detection module to a movable shuttle of a fluorescence analysis apparatus;

an excitation light generator mounted inside the housing such that an excitation optical path from the excitation light generator to the opening has a constant length; and

an emission light detector mounted inside the housing such that a detection optical path from the opening to the emission light detector has a constant length,

wherein when the detection module is attached to the movable shuttle, the opening is oriented toward the plurality of wells and the shuttle is movable to position the detection module in optical communication with different wells of the plurality of wells.

9. 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 detachably attached to a movable shuttle, the detection module including a housing having an opening oriented toward the plurality of wells, an excitation light generator disposed within the housing such that an excitation optical path from the excitation light generator to the opening has a constant length and an emission light detector disposed within the housing such that a detection optical path from the opening to the emission light detector has a constant length;

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 opening is sequentially positioned in optical communication with each of the plurality of sample wells.

10. The method of claim 9 wherein the target molecule is a nucleic acid sequence.

11. The method of claim 10 wherein the reaction is a polymerase chain reaction (PCR).

12. The method of claim 9 wherein the step of scanning includes:

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

briefly activating the excitation light generator;

detecting a fluorescent response using the emission light detector; and

repositioning the detection module such that the opening is in optical communication with a different well of the sample wells.

13. The method of claim 12 wherein during the step of scanning, the detection module continues in motion while the excitation light generator is briefly activated.