METHODS AND SYSTEMS FOR MONITORING MULTIPLE OPTICAL SIGNALS FROM A SINGLE SOURCE

• US 20100331212A1
• Filed: 09/03/2010
• Published: 12/30/2010
• Est. Priority Date: 08/11/2005
• Status: Abandoned Application

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Turner, Stephen, Lundquist, Paul, Zaccarin, Denis
• Application Number:
  US12/875,853
• Publication Number:
  US 20100331212A1
• Time in Patent Office: Days
• US Class Current: 506/12
• CPC Class Codes: G01J 3/2803 using photoelectric array detector
• CPC Class Codes: G01J 3/36 Investigating two or more bands of a spectrum by separate detectors
• 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 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

Methods and systems are described for determining information about template sequences by simultaneously providing excitation over time to a plurality of confined sources on a substrate, each confined source carrying out a template mediated addition of nucleotides with a polymerase enzyme. The nucleotides have different labels, each with a different emission wavelength profile. Optical elements direct optical signals from the different NTPs within each optical confinement to different detector locations. Observing the optical signals over time allows for obtaining sequence information about the template nucleic acids.

Claims

1. An optical analysis method comprising:

simultaneously providing excitation radiation over time to each of a plurality of confined sources on a substrate, wherein each confined source comprises a polymerase enzyme associated with a template nucleic acid sequence;

exposing the polymerase enzymes to polymerase reaction conditions including two or more differently fluorescently labeled types of NTPs, whereby each differently labeled type of NTP has a different emission wavelength profile;

passing the emitted light through an optical train such that for each confined source the optical signal from each type of NTP is directed along a different optical path than each other type of NTP, such that the optical signal from each type of NTP is directed to a different detector location;

observing the optical signals over time from each confined source to determine a time sequence of NTP incorporation, thereby obtaining information about the template nucleic acid sequences.

2. The method of claim 1 wherein the optical signals are detected using a single detector.

3. The method of claim 1 wherein the optical signals are detected using two or more discrete detectors or detector arrays.

4. The method of claim 1 wherein the optical train comprises one or more wedge prism or optical grating to direct the optical signals along different optical paths to different detector locations.

5. The method of claim 1 wherein the confined sources comprises reaction regions.

6. The method of claim 5 wherein the confined reaction regions comprise wells in a multiwell plate.

7. The method of claim 5 wherein confined reaction regions comprise features on a molecular array.

8. The method of claim 5, wherein the confined reactions region comprise zero mode waveguides.

9. The method of claim 1 wherein the polymerase enzyme comprises a DNA polymerase.

10. The method of claim 1 wherein the two or more differently fluorescently labeled types of NTPs comprise four differently fluorescently labeled types on NTPs.

11. The method of claim 10 wherein the four differently fluorescently labeled types on NTPs correspond to A, T, G, and C.

12. A system for performing optical analyses comprising:

a substrate comprising a plurality of confined sources, wherein each confined source comprises a polymerase enzyme associated with a template nucleic acid sequence;

an excitation radiation source that provides excitation radiation to the plurality of confined sources on the substrate, whereby the confined sources are in contact with a solution comprising two or more differently fluorescently labeled types of NTPs, whereby each differently labeled type of NTP has a different emission wavelength profile;

an optical train that directs the optical signal from each type of NTP within each confined source along a different optical path than each other type of NTP, such that the optical signal from each type of NTP is directed to a different detector location, and

one or more detectors that measure the optical signals over time to determine a time sequence of NTP incorporation for the polymerases within each confined source, thereby obtaining information about the template nucleic acid sequence.

13. The system of claim 12 wherein the optical signals are detected using a single detector.

14. The system of claim 12 wherein the optical signals are detected using two or more discrete detectors or detector arrays.

15. The system of claim 12 wherein the optical train comprises one or more wedge prism or optical grating to direct the optical signals along different optical paths to different detector locations.

16. The system of claim 12 wherein the confined sources comprises reaction regions.

17. The method of claim 16 wherein the confined reaction regions comprise wells in a multiwell plate.

18. The system of claim 16 wherein confined reaction regions comprise features on a molecular array.

19. The system of claim 16, wherein the confined reactions region comprise zero mode waveguides.

20. The system of claim 12 wherein the one or more detectors comprise one or more photodiode arrays.

21. The system of claim 12 wherein the one or more detectors comprise one or more charge coupled devices (CCDs).