Methods and systems for simultaneous real-time monitoring of optical signals from multiple sources

• US 8,264,687 B2
• Filed: 05/06/2011
• Issued: 09/11/2012
• Est. Priority Date: 02/13/2006
• Status: Active Grant

• Current Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Original Assignee: Pacific Bioscience of California Incorporated (L'Oreal SA)
• Inventors: Lundquist, Paul, Zaccarin, Denis, Lacroix, Yves, Maxham, Mark, Foquet, Mathieu, Turner, Stephen
• Primary Examiner(s): Geisel, Kara E
• Application Number:
  US13/102,551
• Publication Number:
  US 20110291027A1
• Time in Patent Office: 494 Days
• Field of Search: 356/300, 356317-318, 356/417, 356/301
• US Class Current: 356/417
• CPC Class Codes: G01J 3/02 Details
• CPC Class Codes: G01J 3/0224 using polarising or depolarising elements
• CPC Class Codes: G01J 3/10 Arrangements of light sources specially adapted for spectrometry or colorimetry
• CPC Class Codes: G01J 3/14 using refracting elements, e.g. prisms G01J3/18, G01J3/26 take precedence prisms per se G02B5/04
• CPC Class Codes: G01J 3/2803 using photoelectric array detector
• CPC Class Codes: G01N 2021/6417 Spectrofluorimetric devices
• CPC Class Codes: G01N 21/6445 Measuring fluorescence polarisation
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 2201/126 Microprocessor processing

Abstract

Methods and systems for real-time monitoring of optical signals from arrays of signal sources, and particularly optical signal sources that have spectrally different signal components. Systems include signal source arrays in optical communication with optical trains that direct excitation radiation to and emitted signals from such arrays and image the signals onto detector arrays, from which such signals may be subjected to additional processing.

Claims

1. A method comprising:

providing a substrate comprising a regular array of more than 100 discrete sources of optical signals;

designating an array of subsets of pixels to correspond with the images of the array of the more than 100 discrete sources on a detector as high fidelity pixels and designating another set of pixels on the detector as low fidelity pixels;

then

illuminating the array of discrete sources with an excitation light source, whereby emitted lights emitted from the discrete sources, the emitted light corresponding to signals, from the discrete sources;

directing the emitted light onto the detector;

combining the data from the pixels within each set of high fidelity data for each discrete source; and

processing the combined data to provide information about the signals from the discrete sources.

2. The method of claim 1 wherein data from the low fidelity pixels is combined.

3. The method of claim 2 wherein the data from the low fidelity pixels is discarded.

4. The method of claim 1 wherein the data from the low fidelity pixels is used in determining the background noise level.

5. The method of claim 1 wherein the array of pixels is oriented in rows and columns, and wherein all of the pixels in some of the rows and all of the pixels in in some of the columns are designated as low fidelity pixels.

6. The method of claim 5 wherein the rows or columns range from 2 to 20 pixels wide.

7. The method of claim 1 wherein the images of set of high fidelity pixels corresponds to the central portion of the image of the discrete source.

8. The method of claim 7 wherein the high fidelity signal corresponds to about 25% to 90% of the radius of the image of the discrete source.

9. The method of claim 1 wherein the pixels designated as high fidelity and as low fidelity is adjusted dynamically.

10. The method of claim 1 wherein the designation of high fidelity and low fidelity pixels is adjusted to optimize for different regions on the array.

11. A system comprising:

a substrate comprising a regular array of more than 100 discrete sources of optical signals;

an optical train configured to direct excitation, radiation from an excitation source to the regular array of more than 100 discrete sources, receive emitted optical signals from the array of discrete sources, and image the emitted optical signals onto a detector comprising an array of pixels;

wherein the detector has, for each discrete source, a regular array of subsets of pixels that correspond to expected images of the discrete sources, which are designated as a set of high fidelity pixels, and other pixels on the detector are designated as low fidelity pixels; and

a first processor configured for combining the data from the pixels within each set of high fidelity data for each discrete source; and

a second processor configured for processing the combined data to provide information about the signals from the discrete sources.

12. The system of claim 11 wherein the detector comprises the first processor.

13. The system of claim 11 wherein the detector comprises a CCD, an EMCCD, or an ICCD.

14. The system of claim 11 wherein the first processor and the second processor are on a computer that is separate from the detector.

15. The system of claim 11 wherein the first processor or second processor is configured to combine data from the low fidelity pixels.

16. The system of claim 11 wherein the first processor or second processor is configured to discard data from the low fidelity pixels.

17. The system of claim 11 wherein the first processor or second processor is configured to use the data from the low fidelity pixels to determine the background noise level.

18. The system of claim 11 wherein the array of pixels is oriented in rows and columns, and wherein all of the pixels in some of the rows and all of the pixels in in some of the columns are designated as low fidelity pixels.

19. The system of claim 18 wherein the rows or columns range from 2 to 20 pixels wide.

20. The system of claim 11 wherein the images of set of high fidelity pixels corresponds to the central portion of the image of the discrete source.

21. The system of claim 11 wherein the high fidelity signal corresponds to about 25% to 90% of the radius of the image of the discrete source.