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

• US 8,149,399 B2
• Filed: 05/09/2011
• Issued: 04/03/2012
• 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, Turner, Stephen, Dixon, John
• Primary Examiner(s): Geisel, Kara E
• Application Number:
  US13/103,278
• Publication Number:
  US 20110287981A1
• Time in Patent Office: 330 Days
• Field of Search: 356317-318, 250/458.1
• US Class Current: 356/317
• 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

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 system for monitoring chemical or biochemical reactions in real time comprising:

a substrate comprising a plurality of discrete signal sources, each discrete signal source configured to carry out a chemical or biochemical reaction, wherein fluorescence emission from each discrete signal source changes as the chemical or biochemical reaction progresses, and wherein the plurality of discrete signal sources are arranged in substantially parallel rows;

an excitation source;

an optical train configured to direct excitation radiation to the substrate and to image the fluorescent emissions from the signal sources onto an array detector, whereby the optical train is configured to divide the excitation light into multiple parallel beams, each beam having a line illumination profile, wherein the peaks of the line illumination profile correspond to rows in the array of signal sources.

2. The system of claim 1 wherein the signal sources within a row are spaced apart by first distance and the rows are spaced apart by a second distance, wherein the second distance is greater than three times the first distance.

3. The system of claim 1 wherein the signal sources emit 2, 3, or 4 different fluorescent emission profiles.

4. The system of claim 1 wherein the excitation radiation is divided into 50 or more parallel beams.

5. The system of claim 1 wherein the excitation radiation is divided using a grating or a diffractive element.

6. The system of claim 1 wherein the chemical or biochemical reaction comprises an immunoassay, enzymatic assay, receptor assay, nucleic acid hybridization assay, nucleic acid synthesis reaction, or cellular assay.

7. The system of claim 1 wherein the chemical or biochemical reaction comprises the consumption, production or conversion of a material.

8. The system of claim 1 wherein the chemical or biochemical reaction comprises single molecule analysis.

9. The system of claim 1 wherein the rows are spaced apart on the substrate by a distance that is at least two times the width of a linear illumination profile.

10. The system of claim 1 wherein the signal sources comprise depressions, wells, or zones that are maintained discrete from other regions.

11. The system of claim 1 wherein the signal sources comprise zero mode waveguides.

12. The system of claim 1 wherein the substrate comprises an optically transparent layer, wherein the signal sources are on top of the transparent layer, and wherein the excitation radiation is directed to the signal sources from below, through the transparent layer.

13. The system of claim 1 wherein the substrate comprises greater than 10,000 signal sources.

14. The system of claim 1 wherein the excitation sources comprises a laser.

15. The system of claim 1 comprising multiple excitation sources, each divided into multiple illumination beams.

16. A system for monitoring chemical or biochemical reactions in real time comprising:

a substrate comprising at least a first subset of signal sources positioned in a first substantial linear orientation, and a second subset of signal sources positioned in a second substantially linear orientation that is substantially parallel to the first linear orientation, the signal sources configured to carry out a chemical or biochemical reaction, wherein fluorescence emission from each discrete signal source changes as the chemical or biochemical reaction progresses;

an excitation source;

an optical train configured to direct excitation radiation to the substrate and to image the fluorescent emissions from the signal sources onto an array detector, the excitation radiation comprising at least a first and a second substantially parallel linear illumination profile, wherein the first linear profile illuminates the first subset of signal sources and the second illumination profile simultaneously illuminates the second subset of signal sources.

17. The system of claim 16 wherein the substrate comprises an optically transparent layer, wherein the signal sources are on top of the transparent layer, and wherein the excitation radiation is directed to the signal sources from below, through the transparent layer.

18. The system of claim 16 wherein the substrate comprises greater than 10,000 signal sources.

19. The system of claim 16 wherein the excitation sources comprises a laser.

20. The system of claim 16 comprising multiple excitation sources, each divided into multiple illumination profiles.