Methods and compositions for calibrating chemical array readers

• US 20050244984A1
• Filed: 04/28/2004
• Published: 11/03/2005
• Est. Priority Date: 04/28/2004
• Status: Abandoned Application

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Parker, Russell Alan
• Application Number:
  US10/834,423
• Publication Number:
  US 20050244984A1
• Time in Patent Office: Days
• US Class Current: 436/172
• CPC Class Codes: G01N 21/278 Constitution of standards
• CPC Class Codes: G01N 21/6452 Individual samples arranged in a regular 2D-array, e.g. multiwell plates

Abstract

The invention provides compositions for calibrating a fluorescence reader. The subject compositions generally relate to a solid support coated in a fluorescent agent, where the solid support has minimal local and global nonuniformities of fluorescence. Also provided by the invention are methods of calibrating a scanner using the subject compositions, and kits and programming for performing the subject methods.

Claims

1. A method for making a uniformly fluorescent solid support, comprising:

photobleaching regions of a non-uniformly fluorescent solid support that fluoresce at an intensity above a pre-determined threshold level to produce a uniformly fluorescent solid support.

2. The method of claim 1, wherein said regions of said non-uniformly fluorescent solid support are photobleached by:

a) assessing fluorescence in said regions using a fluorescence reader; and

,

b) photobleaching said regions until they do not exceed said pre-determined threshold.

3. The method of claim 2, wherein said reader is a scanner.

4. The method of claim 1, wherein said region are photobleached using a laser and a light mask.

5. The method of claim 4, wherein said fluorescent solid support is read to provide a data file for said non-uniformly fluorescent solid support, and said light mask is designed using said data file.

6. The method of claim 1, wherein said method comprises:

placing said non-uniformly fluorescent solid support in a reader;

assessing fluorescence of a first region of said non-uniformly fluorescent solid support using said reader;

photobleaching said first region using a laser of said reader until said region has a pre-determined level of fluorescence;

assessing fluorescence of a second region of said non-uniformly fluorescent solid support using said reader; and

,

photobleaching said second region using a laser of said reader until said second region also has said pre-determined level of fluorescence.

7. The method of claim 6, wherein said region is a pixel.

8. The method of claim 6, wherein said assessing and photobleaching steps are repeated until said solid support has a uniform fluorescence.

9. The method of claim 1, wherein said uniformly fluorescent solid support has less than less than about 3% local and global nonuniformity in fluorescence.

15. A method for calibrating a chemical array reader, comprising:

a) reading a composition of claim 9 with said reader to obtain a data file; and

b) calibrating said chemical array reader using said data file.

10. A composition, comprising:

a solid support coated in a fluorescent agent,

wherein said solid support has minimal local and global nonuniformities of fluorescence and contains a plurality of photobleached regions.

11. The composition of claim 10, wherein said solid support comprises a polymer layer comprising said fluorescent agent.

12. The composition of claim 10, wherein said solid support is a planar solid support.

13. The method of claim 10, wherein said solid support has less than less than about 3% local and global nonuniformity.

14. A chemical array reader comprising a composition of claim 10.

16. The method of claim 14, wherein said calibrating comprises:

assessing focus position of said chemical array reader;

assessing dynamic focus of said chemical array reader;

assessing a scanner mirror of said chemical array reader;

or

assessing jitter of said array reader.

17. The method of claim 16, further comprising adjusting said reader.

18. A method for performing a hybridization assay, said method comprising:

(a) calibrating an array reader with a composition of claim 10 to provide a calibrated array reader,

(b) performing a hybridization assay with a chemical array, and

(c) reading said chemical array with said calibrated array reader to provide data.

19. The method of claim 18, wherein said array reader is an array scanner.

20. A method comprising transmitting to a remote location data obtained by a method of claim 18.

21. A method comprising receiving from a remote location data obtained by a method of claim 18.

22. A kit for calibrating a chemical array reader, said kit comprising:

a composition of claim 10.