Assay instrument for detecting optical signals from samples

• US 8,748,789 B2
• Filed: 05/30/2013
• Issued: 06/10/2014
• Est. Priority Date: 03/06/2010
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Triener, Alexander, Allegoren, Erik, Feng, Wenyi, Buermann, Dale, Olson, Erik, Osmus, James
• Primary Examiner(s): LE, QUE TAN
• Application Number:
  US13/905,633
• Publication Number:
  US 20130261028A1
• Time in Patent Office: 376 Days
• Field of Search: 250/201.2, 250/559.4, 250/226, 250/216, 250/239, 250/222.1, 356/246, 356/440, 359361-363, 359385-390, 4352872-2874, 435/7.1, 435 616- 619
• US Class Current: 250/201.2
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6874 involving nucleic acid arrays, e.g. sequencing by hybridisation
• 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 21/6456 Spatial resolved fluorescence measurements; Imaging

Abstract

An optical system configured to detect optical signals during imaging sessions. The optical system includes an objective lens that has a collecting end that is positioned proximate to a sample and configured to receive optical signals therefrom. The optical system also includes a removable path compensator that is configured to be located at an imaging position between the collecting end of the objective lens and the sample. The path compensator adjusts an optical path of the light emissions when in the imaging position. Also, the optical system includes a transfer device that is configured to move the path compensator. The transfer device locates the path compensator at the imaging position for a first imaging session and removes the path compensator from the imaging position for a second imaging session.

Claims

1. An assay instrument for detecting optical signals from samples, comprising:

a sample stage comprising a first interface and a second interface, the first interface comprising a platform for holding a sample on an open-faced support and for imaging of an external surface and the second interface comprising a platform for holding a sample in a flow cell for imaging of at least one internal surface, wherein the sample stage is configured to couple to the flow cell with fluidics connectors;

an objective lens positioned proximate to the sample stage for imaging;

an optical detector for detecting optical signals from the open-faced support and flow cell; and

a moving mechanism for selectively moving one or more optical components into an optical pathway between the sample stage and the optical detector to either a first configuration for surface-imaging of the open-faced support or a second configuration for imaging the flow cell.

2. The assay instrument of claim 1, further comprising a fluidic network that is configured to couple to the flow cell with fluidics connectors, wherein the fluidic network comprises a fluid storage system for storing reagents for sequencing by synthesis.

3. The assay instrument of claim 1, further comprising a protocol module for instructing the assay instrument to perform a sequencing by synthesis protocol for the flow cell and for instructing the assay instrument to perform a microarray scanning protocol for the open-faced support.

4. The assay instrument of claim 3, wherein the protocol module is configured to operate according to priority statuses of the samples whereby lower priority samples are paused when a schedule conflict arises.

5. The assay instrument of claim 4, wherein a sample in a flow cell has a higher priority than a sample on an open-faced support.

6. The assay instrument of claim 1, wherein a working distance exists between the sample stage and the objective lens, the working distance being less than 5000 microns.

7. The assay instrument of claim 1, wherein the resolution of the assay instrument is sufficient to individually resolve features or sites that are separated by a distance of less than 15 μ

m.

8. The assay instrument of claim 1, wherein the objective lens comprises

(i) a collecting end that is configured to be positioned proximate to one of the first and second interfaces on the sample stage and configured to receive optical signals therefrom;

and

(ii) an afocal end configured to transmit the optical signals toward the optical detector, and

wherein the one or more optical components comprise a first removable path compensator for adjusting an optical path of the optical signals when positioned between the collecting end of the objective lens and the sample.

9. The assay instrument of claim 8, wherein the one or more optical components further comprise a second removable path compensator for adjusting the optical path of the optical signals when located at an afocal position with respect to the objective lens.

10. The assay instrument of claim 9, wherein the moving mechanism includes a transfer device located at a fixed position with respect to the sample stage, the transfer device configured to move the first removable path compensator into and out of an imaging position between the collecting end of the objective lens and the sample stage.

11. The assay instrument of claim 10, wherein the transfer device comprises a platform assembly configured to removably couple the first path compensator to the objective lens, a motor assembly configured to move the sample stage in an x-y direction and also along a z-direction to and from the objective lens, and a bridge member configured to couple the platform assembly to the motor assembly.

12. The assay instrument of claim 9, wherein the first removable path compensator is configured to adjust the optical path between a configuration for imaging the flow cell and a configuration for imaging the open-faced support.

13. The assay instrument of claim 12, wherein the second path compensator is configured to adjust the optical path between a configuration for imaging a top surface of the flow cell and a configuration for imaging a bottom surface of the flow cell.

14. The assay instrument of claim 9, wherein the first path compensator is operatively coupled to the objective lens through magnetic forces.

15. The assay instrument of claim 1, wherein the optical signals propagate in a beam direction, and wherein the one or more optical components comprise an optical device which has a rotation axis and comprises first and second bandpass filters, the rotation axis extending in a non-parallel manner with respect to the beam direction, the first and second bandpass filters having fixed orientations with respect to the rotation axis, the optical device being selectively rotatable about the rotation axis to position at least one of the first and second bandpass filters within the optical path between the optical detector and the objective lens along which the optical signals propagate.

16. The assay instrument of claim 1, wherein portions of the optical signals are directed by the one or more optical components along a first optical path and a second optical path between the optical detector and the objective lens;

wherein

the one or more optical components comprise first and second optical devices located proximate to the first and second optical paths, respectively, each of the first and second optical devices having a rotation axis and comprising a plurality of bandpass filters, the bandpass filters having fixed orientations with respect to the rotation axis, wherein each of the first and second optical devices selectively rotates about the corresponding rotation axis to position at least one of the bandpass filters within the corresponding optical path.

17. The assay instrument of claim 1, wherein

the one or more optical components comprise a removable optical wedge, which is positioned in the optical path between the optical detector and the objective lens for detecting first optical signals and removed from the optical path for detecting second optical signals, the optical wedge directing the first optical signals when positioned in the optical path so that the first optical signals are incident upon the sample detector, the second optical signals being incident upon the sample detector when the optical wedge is removed from the optical path.

18. The assay instrument of claim 1, further comprising an excitation light source assembly comprising first and second excitation light sources, the first and second excitation light sources configured to excite the sample during separate first and second imaging sessions, respectively, the sample generating corresponding light emissions when excited by each of the first and second excitation light sources.

19. The assay instrument of claim 18, wherein the one or more optical components comprise a beam splitter separating the corresponding light emissions of each of the first and second excitation light sources into reflected and transmitted portions, the transmitted portions of the first and second excitation light sources being directed along a common transmitted optical path, the reflected portions of the first and second excitation light sources being directed along a common reflected optical path.

20. The assay instrument of claim 19, wherein the one or more optical components include a plurality of optical wedges, each of the transmitted and reflected optical paths having an optical wedge that is selectively moveable, wherein the optical wedge of the reflected optical path redirects the reflected portion during the first imaging session and the optical wedge of the transmitted optical path redirects the transmitted portion during the second imaging session.