Digital image analysis method for enhanced and optimized signals in fluorophore detection

• US 20040060987A1
• Filed: 04/29/2003
• Published: 04/01/2004
• Est. Priority Date: 05/07/2002
• Status: Abandoned Application

• Current Assignee: PriTest Inc.
• Original Assignee: PriTest Inc.
• Inventors: Green, Larry R.
• Application Number:
  US10/425,222
• Publication Number:
  US 20040060987A1
• Time in Patent Office: Days
• US Class Current: 235/491
• CPC Class Codes: B01L 3/545 for laboratory containers
• CPC Class Codes: G06K 19/06009 with optically detectable marking G06K19/063, G06K19/08 take precedence
• CPC Class Codes: G06K 19/06046 Constructional details

Abstract

The present invention concerns methods and apparatus for detecting and/or identifying analytes, using arrays of binding moieties. In preferred embodiments, the arrays are attached to glass slides. Fluorescent signals obtained from the slides are analyzed by a digital image subtraction method. In preferred embodiments, the glass slides are labeled using a binary code that may be used to identify the lot number and/or date of manufacture of the arrays.

Claims

1. A method comprising marking an item with a fluorescent binary code.

2. The method of claim 1, wherein the item is a glass slide, waveguide, chip, protein chip, nucleic acid chip, DNA chip or antibody array.

3. The method of claim 1, wherein the binary code identifies the lot number of the item.

4. The method of claim 1, wherein the binary code identifies the manufacturing date of the item.

5. The method of claim 1, wherein the binary code comprises two rows of spots.

6. The method of claim 1, wherein the first row of spots comprises double spots and the second row of spots comprises single spots.

7. The method of claim 6, wherein the binary code is arranged as disclosed in FIG. 3.

8. The method of claim 5, wherein the two rows of spots further comprise a binary 0 reference spot, a binary 1 reference spot, and an ID reference index spot for the binary code.

9. The method of claim 8, wherein the two rows of spots further comprise an ID check sum validation spot.

10. The method of claim 9, wherein the two rows of spots further comprise fourteen code spots.

11. An item marked by the method of claim 1.

12. A method comprising

a) obtaining a first digital image of one or more spots;

b) obtaining a second digital image of the same one or more spots; and

c) subtracting one image from the other image.

13. The method of claim 12, further comprising analyzing the subtracted data to determine the amount of analyte bound to each spot.

14. The method of claim 12, further comprising averaging two or more pictures to obtain each image.

15. The method of claim 14, further comprising averaging five pictures to obtain each image.

16. The method of claim 12, wherein the images are obtained from an array of binding moieties exposed to excitatory light.

17. The method of claim 16, wherein the light from the array is not filtered.

18. The method of claim 17, wherein the array in the first image comprises primary antibodies and the first image is subtracted from the second image.

19. The method of claim 18, wherein the array in the second image comprises primary antibodies, analytes, biotinylated secondary antibodies and streptavidin conjugated fluorophores.

20. The method of claim 17, wherein the second image is subtracted from the first image and the array in the first image comprises nucleic acid probes, target nucleic acids and fluorophores.

21. The method of claim 20, wherein the array in the second image comprises nucleic acid probes.

22. The method of claim 21, further comprising washing the array to remove target nucleic acids after the first image is obtained.