CONFOCAL IMAGING METHODS AND APPARATUS

• US 20100012825A1
• Filed: 08/06/2009
• Published: 01/21/2010
• Est. Priority Date: 11/23/2005
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Triener, Alexander, Wang, Mark, Che, Diping, Feng, Wenyi, Kain, Robert, Kotseroglou, Theofilos
• Granted Patent: US 7,960,685 B2
• Time in Patent Office: Days
• US Class Current: 250/235
• CPC Class Codes: G01N 2021/6421 Measuring at two or more wavelengths
• 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
• CPC Class Codes: G01N 21/6456 Spatial resolved fluorescence measurements; Imaging
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G02B 21/0032 Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06
• CPC Class Codes: G02B 21/004 fixed arrays, e.g. switchable aperture arrays

Abstract

The invention provides imaging apparatus and methods useful for obtaining a high resolution image of a sample at rapid scan rates. A rectangular detector array having a horizontal dimension that is longer than the vertical dimension can be used along with imaging optics positioned to direct a rectangular image of a portion of a sample to the rectangular detector array. A scanning device can be configured to scan the sample in a scan-axis dimension, wherein the vertical dimension for the rectangular detector array and the shorter of the two rectangular dimensions for the image are in the scan-axis dimension, and wherein the vertical dimension for the rectangular detector array is short enough to achieve confocality in a single axis.

Claims

1. A detection apparatus comprising:

(a) a rectangular detector array comprising a plurality of pixel elements in a horizontal axis and a plurality of pixel elements in a vertical axis, wherein said horizontal axis is longer than said vertical axis; and

(b) optics positioned to direct radiation to said rectangular detector array, wherein said rectangular detector array is configured to collect electrical responses from pixel elements in the vertical axis in a common bin, thereby achieving confocality in a single axis of said detector, wherein said single axis is said vertical axis.

2. The apparatus of claim 1, further comprising a radiation source positioned to send excitation radiation to at least a portion of a sample region, wherein said optics are positioned to direct said radiation from said at least a portion of said sample region to said rectangular detector array.

3. The apparatus of claim 2, further comprising a line generator positioned to receive excitation radiation from said radiation source and to send a radiation line to said at least a portion of said sample region.

4. The apparatus of claim 3, wherein the ratio of the shorter of two rectangular dimensions for said radiation line to the Rayleigh resolution of the imaging optics is in the range of 0.1 to 10.

5. The apparatus of claim 2, further comprising a scanning device configured to scan said at least a portion of said sample region in a scan-axis dimension, wherein the vertical axis for said rectangular detector array is in said scan-axis dimension.

6. The apparatus of claim 5, wherein said optics are positioned to direct said radiation to said rectangular detector array as a rectangular image of said at least a portion of said sample region, and wherein the shorter of the two rectangular dimensions for said image correlates with the vertical axis of said rectangular array.

7. The apparatus of claim 6, wherein said shorter of the two rectangular dimensions for said image is in said scan axis dimension.

8. The apparatus of claim 1, wherein the aspect ratio of said rectangular detector array is greater than 20.

9. The apparatus of claim 1, wherein said apparatus is configured to obtain an image at a Rayleigh resolution between 0.2 and 10 micrometers.

10. The apparatus of claim 1, wherein said rectangular detector array is further configured to collect electrical responses from pixel elements in the horizontal axis in a common bin.

11. A method of detecting a sample, comprising

(a) directing radiation from at least a portion of a sample region to a rectangular detector array, wherein said rectangular detector array comprises a plurality of pixel elements in a horizontal axis and a plurality of pixel elements in a vertical axis, wherein said horizontal axis is longer than said vertical axis;

(b) collecting electrical responses from a plurality of pixel elements of the vertical axis in a common bin; and

(c) reading the collected electrical responses from said common bin as a single value, thereby achieving confocality in a single axis of said detector, wherein said single axis is said vertical axis.

12. The method of claim 11, wherein said radiation from said at least a portion of said sample is produced by contacting said at least a portion of said sample with excitation radiation.

13. The method of claim 12, wherein said excitation radiation comprises a radiation line.

14. The method of claim 13, wherein the ratio of the shorter of two rectangular dimensions for said radiation line to the Rayleigh resolution of the imaging apparatus is in the range of 0.1 to 10.

15. The method of claim 11, further comprising scanning said at least a portion of said sample region in a scan-axis dimension, wherein the vertical axis for said rectangular detector array is in said scan-axis dimension.

16. The method of claim 15, wherein said radiation is directed to said rectangular detector array as a rectangular image of said at least a portion of said sample region, and wherein the shorter of the two rectangular dimensions for said image correlates with the vertical axis of said rectangular array.

17. The method of claim 16, wherein said shorter of the two rectangular dimensions for said image is in said scan axis dimension.

18. The method of claim 11, wherein said radiation from said at least a portion of said sample forms a rectangular image of said at least a portion of said sample.

19. The method of claim 11, wherein an image of said at least a portion of said sample region is obtained at a Rayleigh resolution between 0.2 and 10 micrometers.

20. The method of claim 11, wherein said sample comprises a microarray.