Instrument for monitoring polymerase chain reaction of DNA

• US 20070148761A1
• Filed: 02/13/2007
• Published: 06/28/2007
• Est. Priority Date: 05/16/1998
• Status: Abandoned Application

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Atwood, John, Lakatos, Edward, Young, Eugene, Stone, Susan, Cerrone, Anthony, Gambini, Michael, Atwood, Judith
• Application Number:
  US11/705,856
• Publication Number:
  US 20070148761A1
• Time in Patent Office: Days
• US Class Current: 435/287.200
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0663 Whole sensors
• CPC Class Codes: B01L 2300/1811 using electromagnetic induction heating
• CPC Class Codes: B01L 3/5085 for multiple samples, e.g. microtitration plates
• 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: C12Q 2545/101 with an internal standard/control
• CPC Class Codes: C12Q 2563/107 fluorescence
• 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 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 2021/6441 with two or more labels
• CPC Class Codes: G01N 2021/6463 Optics
• CPC Class Codes: G01N 2021/6482 Sample cells, cuvettes
• CPC Class Codes: G01N 21/274 Calibration, base line adjustment, drift correction
• 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
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 2201/0245 with insertable-removable part
• CPC Class Codes: G01N 2201/06113 : Coherent sources; lasers
• CPC Class Codes: G01N 2201/0628 : Organic LED [OLED]
• CPC Class Codes: G01N 2201/0635 : Structured illumination, e.g. with grating
• CPC Class Codes: G01N 2201/0636 : Reflectors
• CPC Class Codes: G01N 2201/068 : Optics, miscellaneous

Abstract

An optical instrument monitors PCR replication of DNA in a reaction apparatus having vials of reaction ingredients. A beam splitter passes an excitation beam to the vials. An emission beam from the reaction ingredients can be passed by the beam splitter to a detector from which a processor can compute DNA concentration. A reference strip with a plurality of reference emitters can be provided which emits reference beams of different intensity, from which the processor can select an optimum emitter for compensating for drift. Exposure time can be automatically adjusted for keeping within optimum dynamic ranges of the detector and processor. A module of the beam splitter and associated optical filters can be associated with a selected dye, and can be replaceable for different dyes.

Claims

1. An instrument comprising:

a plurality of spaced-apart vials;

a light source adapted to direct an excitation beam along an excitation beam path simultaneously toward each of the plurality of vials;

a beam splitter disposed along the excitation beam path between the light source and the plurality of vials;

a lens disposed along the excitation beam path between the light source and the plurality of vials; and

a detector disposed along an emission beam path and arranged to receive emission beams emitted from the plurality of vials and through the lens;

wherein the lens and the beam splitter are disposed along an emission beam path between the plurality of vials and the detector, and the beam splitter is arranged to split apart the excitation beam path and the emission beam path.

2. The instrument of claim 1, wherein the detector generates primary data signals representative of the emission beams.

3. The instrument of claim 2, further comprising a processor to receive the primary data signals and compute corresponding concentrations in the respective vials.

4. The instrument of claim 1, further comprising a detector lens disposed along an emission beam path, between the lens and the detector.

5. The instrument of claim 1, further comprising an excitation beam filter disposed along an excitation beam path between the light source and the lens.

6. The instrument of claim 1, further comprising an emission beam filter disposed along an emission beam path between the lens and the detector.

7. The instrument of claim 1, further comprising a fold mirror disposed along the excitation beam path, between the light source and the plurality of vials.

8. The instrument of claim 1, further comprising a plurality of focusing lenses disposed along excitation beam paths between the lens and the plurality of vials.

9. The instrument of claim 1, wherein the instrument is adapted to simultaneously direct a plurality of excitation beams respectively toward the plurality of vials.

10. The instrument of claim 1, further comprising reaction ingredients disposed in each of the plurality of vials.

11. The instrument of claim 10, wherein the reaction ingredients include a concentration of a marker dye that emits emission beams when illuminated with an excitation beam.

12. The instrument of claim 10, wherein the reaction ingredients include components for polymerase chain reaction.

13. The instrument of claim 1, wherein the plurality of vials comprises 96 vials.

14. The instrument of claim 1, wherein the plurality of vials comprises a plastic unitary tray.

15. The instrument of claim 1, wherein the plurality of vials comprises 96 vials in a plastic unitary tray.

16. A method comprising:

directing an excitation beam along an excitation beam path toward each of a plurality of vials, simultaneously;

splitting the excitation beam into light that is reflected and light that is passed;

directing light that is reflected from the beam splitter through a lens disposed along the excitation beam path between the light source and the plurality of vials, and simultaneously into each of the plurality of vials, to cause the plurality of vials to emit emission beams;

directing the emission beams from the plurality of vials through the beam splitter and to a detector; and

detecting the emission beams emitted from the plurality of vials.

17. The method of claim 16, wherein the wells contain different respective samples.

18. The method of claim 16, wherein the different respective samples each include a concentration of a marker dye that emits emission beams in response to being illuminated by the excitation beam, and wherein the method further includes generating primary data signals representative of the detected emission beams.

19. The method of claim 18, further comprising processing the primary data signals to compute corresponding concentrations of marker dye in the respective wells.

20. The method of claim 16, wherein the directing the emission beams from the plurality of vials through the beam splitter and to a detector, comprises directing the emission beams through a lens disposed along an emission beam path between the plurality of vials and the beam splitter.