Data processing system and methods

• US 9,530,207 B2
• Filed: 01/29/2015
• Issued: 12/27/2016
• Est. Priority Date: 01/13/2010
• Status: Active Grant

• Current Assignee: Illumina Incorporated
• Original Assignee: Illumina Incorporated
• Inventors: Garcia, Francisco, Maisinger, Klaus, Tanner, Stephen, Moon, John, Mann, Tobias, Parkinson, Michael Lawrence, Cox, Anthony James, Ge, Haifang H.
• Primary Examiner(s): BEKELE, MEKONEN T
• Application Number:
  US14/608,471
• Publication Number:
  US 20150243028A1
• Time in Patent Office: 698 Days
• Field of Search: 382/129, 382/128, 382/209, 382/225, 382/294
• US Class Current: 1/1
• CPC Class Codes: G06T 2207/10024 Color image
• CPC Class Codes: G06T 2207/30024 Cell structures in vitro; Tissue sections in vitro
• CPC Class Codes: G06T 2207/30072 Microarray; Biochip, DNA array; Well plate
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06T 7/0014 using an image reference approach
• CPC Class Codes: G06T 7/246 using feature-based methods, e.g. the tracking of corners or segments
• CPC Class Codes: G06T 7/248 involving reference images or patches
• CPC Class Codes: G06T 7/33 using feature-based methods
• CPC Class Codes: G06T 7/337 involving reference images or patches
• CPC Class Codes: G06T 7/90 Determination of colour characteristics

Abstract

Methods and systems for analysis of image data generated from various reference points. Particularly, the methods and systems provided are useful for real time analysis of image and sequence data generated during DNA sequencing methodologies.

Claims

1. A method of tracking the location of features of a specimen across a set of images of the specimen captured at different reference points, the set of images comprising a plurality of cycles of images, each cycle comprising different images, each image comprising signals having a single color, wherein the signal color is different for each of the different images, said method comprising the steps of:

(a) selecting a subset of images, wherein the images of the subset depict signals corresponding to features of the specimen, and wherein the images of the subset are captured at different reference points;

(b) selecting a primary image from the subset of images, comprising;

(i) selecting the different color images from a single cycle within the subset of images,

(ii) merging signals from at least two different color images of each cycle into a candidate primary image for said cycle,

(iii) repeating steps (i) and (ii), thereby forming a plurality of candidate primary images, and

(iv) selecting the candidate primary image from among the candidate primary images that includes the most signals, thereby obtaining a primary image;

(c) registering the signals depicted in the images of the subset of images with the signals depicted in the primary image, so as to determine the location of the signals depicted in the images with respect to each other, thereby producing signal clumps;

(d) selecting a signal from each of said signal clumps, thereby forming a template that permits the identification of the locations of features of said specimen; and

(e) registering remaining images in said set of images with said template.

2. The method of claim 1, wherein each cycle comprises four different images.

3. The method of claim 1, wherein said specimen comprises at least one tile comprising an array of molecules.

4. The method of claim 3, wherein said specimen comprises a plurality of tiles.

5. The method of claim 3, wherein said array of molecules comprises a plurality of features.

6. The method of claim 5, wherein a feature of said plurality of features comprises multiple copies of a molecule of said array of molecules.

7. The method of claim 6, wherein said molecule comprises a nucleic acid.

8. The method of claim 7, further comprising providing a labeled nucleotide base to said array of molecules, thereby extending a primer hybridized to a nucleic acid within a feature so as to produce a signal corresponding to a feature comprising said nucleic acid.

9. The method of claim 8, wherein the different color corresponds to a different nucleotide base.

10. The method of claim 9, wherein each image of the set of images comprises signals having a single color selected from at least four different colors.

11. The method of claim 10, wherein four different labeled nucleotide bases are provided to said array of molecules so as to produce four different images, thereby producing a cycle of four color images that corresponds to the four possible nucleotides present at a particular position in said nucleic acid.

12. The method of claim 11, wherein said primary image comprises either a single image or a compilation of multiple merged images.

13. The method of claim 1, further comprising the step of selecting a secondary image, wherein said secondary image is the candidate primary image that includes the second most signals.

14. The method of claim 13, wherein said secondary image comprises either a single image or a compilation of multiple merged images.

15. The method of claim 14, further comprising registering the signals from the secondary image with the signals from the primary image.

16. The method of claim 15, further comprising registering signals from nonselected images with the signals from the secondary image, wherein said nonselected images are obtained from the image cycle from which the primary image was obtained.

17. The method of claim 16, further comprising registering signals from remaining images in the subset with signals from the primary image.

18. The method of claim 1, wherein the signals from at least two different color images are subject to cross-talk, and wherein cross-talk between two signal channels allows signals from one color image to appear in the other color image.

19. The method of claim 9, wherein the signals from at least two different color images comprise signals corresponding to an A nucleotide base and a C nucleotide base.

20. The method of claim 1, wherein said template that permits the identification of the locations of features of said specimen is saved to a locations file.