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

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.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic illustration of an overall system the present invention.

FIG. 2 provides a schematic illustration of an array of signal sources on a substrate, such as zero mode waveguides.

FIG. 3 illustrates an alternative configuration of signal sources to accommodate signal separation detection systems of the invention.

FIG. 4 schematically illustrates the separated imaged signals derived from the substrate shown in FIG. 3, providing an overlay of the signal sources from that substrate on the imaged signals.

FIG. 5 schematically illustrates the substrate and optical train of the systems of the invention that includes optical componentry for the separation and detection of spectrally resolvable signal components.

FIG. 6 provides a schematic illustration of a system of the present invention that includes optical componentry for simultaneous illumination of larger numbers of signal sources on the substrates.

FIG. 7 provides an example of an optical train employing a deflective element in dividing one illumination beam into multiple beams.

FIG. 8 provides an alternative example of use of a deflective element in dividing illumination beams.

FIG. 9 schematically illustrates an optical train employing conventional beam splitting optics to generate multiple illumination beams for subsequent linearization.

FIG. 10 schematically illustrates an optical train that utilizes polarizing beam splitters in producing multiple illumination beams and/or lines.

FIG. 11 schematically illustrates a number of alternative optical configurations for splitting and directing different polar beam components of an illumination beam.

FIG. 12 schematically illustrates an optical train that multiplexes the beam splitting function of the optical trains shown in, e.g., FIGS. 11 and 12.

FIG. 13 provides a comparative plot of illumination profiles of flood illumination and multi-line illumination incident upon a substrate.

Claims

1. An analytical device, comprising: a substrate;a plurality of signal sources disposed upon the substrate, the signal sources being arrayed upon the substrate in a plurality of substantially parallel rows, each of the plurality of parallel rows comprising a plurality of signal sources, and wherein two adjacent signal sources in a row are spaced apart by a first distance, and wherein two adjacent rows of signal sources are spaced apart by a second distance, wherein the second distance is at least three times greater than the first distance.

2. The analytical device of claim 1, wherein each of the plurality of signal sources on the substrate comprises a well disposed in a surface of the substrate.

3. The analytical device of claim 2, wherein the substrate comprises a transparent layer and an opraque layer, and each well comprises an aperture disposed through the opaque layer to the transparent layer.

4. The analytrical device of claim 3, wherein each well comprises a zero mode waveguide.

5. The analytical device of claim 1, wherein the first distance is from about 100 nm to about 1 mm and the second distance is from about 200 nm to about 10 mm.

6. The analytical device of claim 1, wherein the substrate comprises at least t 10 substantially parallel rows of signal sources.

7. The analytical device of claim 1, wherein the substrate comprises at least 100 substantially parallel rows of signal sources.

8. An analytical system, comprising: a substrate having a plurality of discrete signal sources disposed thereon;an excitation light source;an optical train positioned to transmit excitation light from the excitation light source to the substrate to illuminate a first plurality of illuminated signal sources and a second plurality of illuminated signal sources, and image signals from the pluarality of illuminated signal sources onto an array detector;wherein one or more of the substrate and the optical train are configured such that the first and second plurality of illuminated signal sources is spaced from the other by a first distance of that is at least three times a cross sectional dimension of an image of a signal from a signal source imaged onto the array detector.

9. The analytical system of claim 8, wherein the substrate comprises a first and second plurality of signal sources adjacent to each other on the substrate and separated by the first distance, wherein the first and second plurality of signal sources corresponds to the first and second plurality of illuminated signal sources.

10. The analytical system of claim 9, wherein the first plurality of signal sources comprises a first row of signal sources and the second plurality of signal sources comprises a second row of signal sources, wherein the first and second rows of signal sources are substantially parallel and separated by the first distance.

11. The analytical system of claim 8, wherein the optical train is configured to provide a first and second illumination profile upon the substrate, wherein the first and second illumination profiles on the substrate are spaced apart by the first distance.

12. A method of analyzing a plurality of signal sources, comprising: providing a substrate having a plurality of discrete signal sources disposed thereon;illuminating a first plurality of illuminated signal sources and a second plurality of illuminated signal sources, wherein the first plurality of illuminated signal sources are spaced apart from the second plurality of illuminated signal sources by a distance that is greater than three times a cross sectional dimension of an image of the signal source imaged on a detector array; andimaging the signal source on a detector array.

13. The method of claim 12, wherein the first plurality of signal sources and the second plurality of signal sources are adjacent to each other and spaced apart by the first distance.

14. The method of claim 13, wherein the illumination step comprises illuminating first and second portions of the substrate that are separated by the first distance upon the substrate by a non-illuminated portion of the substrate, the first plurality of illuminated signal sources being disposed in the first portion of the substrate and the second plurality of illuminated signal sources being disposed in the second portion of the substrate.

15. The method of claim 13, wherein the substrate is flood illuminated and the first and second plurality of illuminated signal sources are spaced apart on the substrate by the first distance.

16. The method of claim 13, wherein the first plurality of signal sources is illuminated with a first illumination beam, and the second plurality of signal sources is illuminated with a second illumination beam.

17. The method of claim 16, wherein the first and second illumination beams each comprise a linear illumination profile.

18. An analytical system, comprising: a substrate having a plurality of signal sources disposed thereon;an excitation light source;an optical train configured to: receive excitation light from the excitation light source;direct it onto the substrate in at least first and second substantially parallel linear illumination profiles, wherein the first and second linear illumination profiles are spaced apart on the substrate by a distance that is at least two times a width of the first linear illumination profile; andreceive optical signals from the substrate an image the oiptical signals onto a detector array.

19. An analytical system, comprising: a substrate comprising a plurality of discrete signal sources disposed thereon, wherein at least a first subset of signal sources are positioned in a first substantial linear orientation, and a second subset of signal sources are positioned in a second substantially linear orientation that is substantially parallel to the first linear orientation;an light source;an optical train for directing light from the light source to the substrate in at least first and second substantially parallel linear illumination profiles, the first linear profile illuminating the first subset of signal sources and the second illumination profile illuminating the second subset of signal sources.

20. The analytical system of claim 19, wherein a position of the first linear illumination profile on the substrate is independently adjustable from a position of the second linear illumination profile on the substrate.

21. The analytical system of claim 19, wherein the optical train comprises at least a first polarizing beam splitter for dividing light from the light source into at least first and second beams, and linearization optics for converting the first and second beams to the first and second linear illumination profiles on the substrate.

22. A method of detecting fluorescent signals from a plurality of signal sources on a substrate, comprising directing excitation radiation at portions of the substrate occupied the plurality of signal sources on a substrate while not directing excitation radiation at portions of the substrate not occupied by the signal sources.

23. A system, comprising: a substrate comprising a plurality of discrete signal sources;an excitation light source;an optical train, positioned to receive excitation light from the excitation light source and direct the excitation light to the substrate, wherein the optical train is configured to direct excitation light in a substantially linear illumination profile at a plurality of signal sources, simultaneously, and simultaneously receive optical signals from the plurality of signal sources and direct the optical signals upon an imaging detector, to detect the optical signals from the plurality of signal sources.

24. A system, comprising: a substrate having at least first and second rows of signal sources disposed thereon;an excitation light source; andan optical train, positioned to receive excitation light from the excitation light source and direct the excitation light to the substrate, wherein the optical train is configured to divide the excitation light into at least first and second discrete beams, and direct each of the at least first and second discrete beams in a substantially linear illumination profile at the substrate, wherein the first beam simultaneously illuminates a plurality of signal sources in the first row of signal sources, and the second beam simultaneously illuminates a plurality of signal sources in the second row of signal sources.

25. A system, comprising: an excitation light source;an optical train positioned to receive excitation light from the excitation light source and direct excitation light to the substrate, the optical train comprising: a polarizing beam splitter to split the excitation light into at least first and second polar component beams; andoptical components for directing each of the first and second polar component beams to different locations on the substrate.

26. A method of analyzing a plurality of signal sources on a substrate, comprising: providing at least first and second adjacent signal sources on a substrate;selectively directing excitation radiation at the first and second signal sources while not substantially illuminating space between the first and second signal sources.

27. The method of claim 26, wherein the first signal source is provided in a first plurality of signal sources, and the second signal source is provided in a second plurality of signal sources, wherein the first and second plurality of signal sources are adjacent to each other, and the step of selectively directing excitation radiation comprises selectively directing excitation radiation at the first and second plurality of signal sources while not substantially illuminating space between the first and second plurality of signal sources.

28. The method of claim 26, wherein the step of selectively directing excitation radiation at the first and second signal sources comprises directing excitation light in first and second separate linear illumination profiles upon the substrate wherein the first linear illumination profile illuminates the first signal source and the second linear illumination profile illuminates the second signal source.