Instrument for monitoring polymerase chain reaction of DNA

• US 7,008,789 B2
• Filed: 08/09/2002
• Issued: 03/07/2006
• Est. Priority Date: 05/16/1998
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Applera Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Young, Eugene F., Gambini, Michael R., Cerrone, Anthony L., Lakatos, Edward J.
• Primary Examiner(s): Redding, David A
• Application Number:
  US10/216,620
• Publication Number:
  US 20020192808A1
• Time in Patent Office: 1,306 Days
• Field of Search: 435/287.2, 435/288.4, 435/288.7, 422/82.08, 378/42, 378/45, 378/48, 250/458.1, 250/483.1, 250/459.1, 250/462.1, 250/461.2, 356/73, 356/141, 356/146
• US Class Current: 435/287.2
• 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 a temperature cycled block with vials of reaction ingredients including dye that fluoresces in presence of double-stranded DNA. A beam splitter passes an excitation beam to the vials to fluoresce the dye. An emission beam from the dye is passed by the beam splitter to a CCD detector from which a processor computes DNA concentration. A reference strip with a plurality of reference emitters emit reference beams of different intensity, from which the processor selects an optimum emitter for compensating for drift. Exposure time is automatically adjusted for keeping within optimum dynamic ranges of the CCD and processor. A module of the beam splitter and associated optical filters is associated with selected dye, and is replaceable for different dyes.

Claims

1. An instrument comprising:

a plurality of spaced-apart reaction ingredient containers;

a light source adapted to direct an excitation beam toward the plurality of containers;

a Fresnel lens disposed along an excitation beam path, between the light source and the plurality of containers;

a detector disposed along an emission beam path and arranged to receive emission beams from the Fresnel lens; and

reaction ingredients disposed in each of the plurality of containers;

wherein the Fresnel lens is also disposed along an emission beam path between the plurality of containers and the detector, the Fresnel lens is adapted to direct emission beams from two or more of the plurality of containers to the detector, and the reaction ingredients include components for polymerase chain reaction.

2. An instrument comprising:

a plurality of spaced-apart reaction ingredient containers;

a light source adapted to direct an excitation beam toward the plurality of containers;

a Fresnel lens disposed along an excitation beam path, between the light source and the plurality of containers; and

a detector disposed along an emission beam path and arranged to receive emission beams from the Fresnel lens;

wherein the Fresnel lens is also disposed along an emission beam path between the plurality of containers and the detector, the Fresnel lens is adapted to direct emission beams from two or more of the plurality of containers to the detector, and the Fresnel lens comprises two or more lenses.

3. An instrument comprising:

a plurality of spaced-apart reaction ingredient containers;

a light source adapted to direct an excitation beam toward the plurality of containers;

a Fresnel lens disposed along an excitation beam path, between the light source and the plurality of containers;

a detector disposed along an emission beam path and arranged to receive emission beams from the Fresnel lens; and

reaction ingredients disposed in each of the plurality of containers;

wherein the Fresnel lens is also disposed along an emission beam path between the plurality of containers and the detector, the Fresnel lens is adapted to direct emission beams from more than one of the plurality of containers simultaneously, and the reaction ingredients include components for polymerase chain reaction.

4. An instrument comprising:

a plurality of spaced-apart reaction ingredient containers;

a light source adapted to direct an excitation beam toward the plurality of containers;

a Fresnel lens disposed along an excitation beam path, between the light source and the plurality of containers; and

a detector disposed along an emission beam path and arranged to receive emission beams from the Fresnel lens;

wherein the Fresnel lens is also disposed along an emission beam path between the plurality of containers and the detector, the Fresnel lens is adapted to direct emission beams from more than one of the plurality of containers simultaneously, and the Fresnel lens comprises two or more lenses.