Optical instrument comprising multi-notch beam splitter

• US 9,823,195 B2
• Filed: 01/23/2013
• Issued: 11/21/2017
• Est. Priority Date: 05/16/1998
• Status: Expired due to Fees

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Inventors: Oldham, Mark F., Young, Eugene F.
• Primary Examiner(s): Bowers, Nathan
• Application Number:
  US13/748,508
• Publication Number:
  US 20130143310A1
• Time in Patent Office: 1,763 Days
• 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 2545/101 with an internal standard/control
• 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 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: G01N 2201/0245 with insertable-removable part
• CPC Class Codes: G01N 2201/0628 Organic LED [OLED]
• CPC Class Codes: G01N 2201/0635 Structured illumination, e.g. with grating
• 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

Abstract

An instrument is provided that can monitor nucleic acid sequence amplification reactions, for example, PCR amplification of DNA and DNA fragments. The instrument includes a multi-notch filter disposed along one or both of an excitation beam path and an emission beam path. Methods are also provided for monitoring nucleic acid sequence amplifications using an instrument that includes a multi-notch filter disposed along a beam path.

Claims

1. An imaging optical instrument, comprising:

an apparatus having a surface configured to hold a two-dimensional array of spaced-apart reaction regions;

an excitation light source comprising at least two, individually addressable light elements, the excitation light source configured to provide at least two excitation beams having different excitation wavelength ranges;

a beam splitter comprising a dichroic beam splitter having two passbands, the beam splitter configured to direct the excitation beams to produce emission beams comprising at least two different emission wavelength ranges;

a detector;

an imaging system comprising a lens configured to direct the excitation beams to simultaneously illuminate the two-dimensional array of spaced-apart reaction regions and to direct the emission beams onto the detector to produce two-dimensional images of the spaced-apart reaction regions;

wherein the beam splitter is configured to reflect light from each of the light elements and to transmit light from the emission beams through the beam splitter and to the detector.

2. The optical instrument of claim 1, further comprising an excitation filter disposed along an excitation path between the excitation light source and the beam splitter, the excitation filter comprising a multiple bandpass filter and configured to transmit light within the different excitation wavelength ranges.

3. The instrument of claim 1, further comprising a Fresnel lens disposed along an excitation path from the excitation light source to the surface and along an emission path from the surface to the detector.

4. The instrument of claim 1, further comprising a mirror disposed along an excitation path from the excitation light source to the surface.

5. The instrument of claim 1, further comprising a plurality of spaced-apart reaction regions disposed on or near the surface, at least some of the reaction regions comprising at least one respective sample including a fluorescent compound configured to produce an emission beam in response to at least one of the excitation beams.

6. The instrument of claim 5, wherein the fluorescent compound comprises a dye, marker, or label.

7. The instrument of claim 5, further comprising a plurality of detectors to detect the emission beams from the reaction regions.

8. The instrument of claim 1, wherein the beam splitter comprises a Rugate filter.

9. The instrument of claim 1, wherein the beam splitter is a dichroic beam splitter.

10. The instrument of claim 1, further comprising a processing device, operatively connected to the detector, the processing device configured to adjust exposure conditions of the detector to maintain a data signal from the detector within a predetermined operating range.

11. The instrument of claim 1, wherein the excitation light source comprises at least two lasers.

12. The instrument of claim 1, wherein the at least two light elements comprises at least two light emitting diodes (“

LEDs”

).

13. The optical instrument of claim 12, wherein the at least two LEDs draw about 10 watts of energy or more.

14. The instrument of claim 1, wherein the detector comprises a charge coupled device (CCD) or a charge injection device (CID).

15. The instrument of claim 1, wherein the detector comprises a plurality of photodiode detectors.

16. The instrument of claim 1, wherein the at least two light elements comprises a plurality of light emitting diodes having different wavelength ranges, the beam splitter being configured to reflect light from each of the light emitting diodes and transmit light from the emission beams.

17. The instrument of claim 1, wherein the detector is a single array detector configured to receive emission beams from more than one of the reaction regions.

18. The optical instrument of claim 1, further comprising a thermal controller for controlling a temperature of a block.

19. An imaging optical instrument, comprising:

an apparatus having a surface configured to hold a two-dimensional array of spaced-apart reaction regions;

an excitation light source comprising at least two, individually addressable light elements, the excitation light source configured to provide at least two excitation beams having different excitation wavelength ranges and to direct the excitation beams to simultaneously illuminate more than one of the reaction regions so as to produce a plurality of emission beams;

a beam splitter comprising a dichroic beam splitter having at least two passbands corresponding to the different excitation wavelength ranges;

a detector;

an imaging system comprising a lens configured to direct the excitation beams to simultaneously illuminate the two-dimensional array of spaced-apart reaction regions and to direct the emission beams onto the detector to produce two-dimensional images of the spaced-apart reaction regions;

wherein the beam splitter is (a) configured to transmit light from each of the light elements through the beam splitter and to reflect light within each of the at least two different emission wavelength ranges or (b) configured to transmit the at least two excitation beams having the different excitation wavelength ranges through the beam splitter and to reflect light within each of the at least two different emission wavelength ranges.

20. An optical instrument, comprising:

an apparatus having a region configured to hold a two-dimensional array of spaced-apart reaction regions;

an excitation light source comprising at least two, independently activated light elements, the excitation light source configured to provide at least two excitation beams having different colors and to direct the excitation beams to simultaneously illuminate more than one of the reaction regions;

a beam splitter comprising a dichroic beam splitter having two passbands, the beam splitter configured to direct the excitation beams to the more than one reaction regions so as to provide emission beams comprising at least two different colors;

a detector;

an imaging system comprising a lens configured to direct the excitation beams to simultaneously illuminate the two-dimensional array of spaced-apart reaction regions and to direct the emission beams onto the detector to produce two-dimensional images of the spaced-apart reaction regions;

wherein the beam splitter is configured to reflect light from each of the light elements and to transmit light from the emission beams through the beam splitter and to the detector.

21. The optical instrument of claim 20, further comprising an excitation filter disposed along an excitation path between the excitation light source and the beam splitter, the excitation filter comprising a multiple bandpass filter and configured to transmit light within the different excitation colors.

22. The instrument of claim 20, further comprising a plurality of spaced-apart reaction regions disposed within the region of the apparatus, at least some of the reaction regions comprising at least one respective sample including a fluorescent compound configured to produce an emission beam in response to at least one of the excitation beams.

23. The instrument of claim 20, wherein the beam splitter is a dichroic beam splitter.

24. The instrument of claim 20, wherein the at least two light elements comprises at least two light emitting diodes (“

LEDs”

).

25. The instrument of claim 20, wherein the detector is a single array detector configured to receive emission beams from more than one of the reaction regions.

26. An optical instrument, comprising:

an apparatus having a region configured to hold a two-dimensional array of spaced-apart reaction regions;

an excitation light source comprising a first light element configured to provide a first excitation beam having first excitation wavelength range and a second light element configured to provide a second excitation beam having second excitation wavelength range that is different from the first excitation wavelength range;

a beam splitter comprising a dichroic beam splitter having two passbands, the beam splitter configured to reflect or transmit the excitation beams to the region of the apparatus to simultaneously illuminate more than one of reaction regions so as to produce emission beams comprising at least two different emission wavelength ranges;

a detector;

an imaging system comprising a lens configured to direct the excitation beams to simultaneously illuminate the two-dimensional array of spaced-apart reaction regions and to direct the emission beams onto the detector to produce two-dimensional images of the spaced-apart reaction regions;

wherein the beam splitter is configured to reflect light from each of the light elements and to transmit light from the emission beams through the beam splitter and to the detector.