Quantified fluorescence microscopy

• US 20070012885A1
• Filed: 08/28/2006
• Published: 01/18/2007
• Est. Priority Date: 02/09/2000
• Status: Abandoned Application

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Montagu, Jean
• Application Number:
  US11/510,631
• Publication Number:
  US 20070012885A1
• Time in Patent Office: Days
• US Class Current: 250/458.100
• CPC Class Codes: B01J 2219/00364 Pipettes
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00691 using robots
• CPC Class Codes: B01L 3/0244 using pins
• CPC Class Codes: B01L 3/0251 Pin and ring type or pin in tube type dispenser
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 2035/1034 Transferring microquantities of liquid
• CPC Class Codes: G01N 2035/1037 Using surface tension, e.g. pins or wires
• CPC Class Codes: G01N 21/274 Calibration, base line adjustment, drift correction
• CPC Class Codes: G01N 21/278 Constitution of standards
• CPC Class Codes: G01N 21/64 Fluorescence; Phosphorescence
• CPC Class Codes: G01N 21/6452 : Individual samples arranged in a regular 2D-array, e.g. multiwell plates
• CPC Class Codes: G01N 21/6458 : Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 35/1065 : Multiple transfer devices
• CPC Class Codes: G02B 21/16 : adapted for ultraviolet illumination ; Fluorescence microscopes G02B21/0076 takes precedence
• CPC Class Codes: G02B 21/34 : Microscope slides, e.g. mounting specimens on microscope slides
• CPC Class Codes: Y10T 428/24355 : Continuous and nonuniform or irregular surface on layer or component [e.g., roofing, etc.]

Abstract

The present invention relates to calibration apparatuses, methods or tools used in microscopy A calibration tool for fluorescent microscopy includes a support, a solid surface layer including a fluorescent material, and a thin opaque mask of non-fluorescent material defining reference feature openings having selected dimensions exposing portions of the surface layer A first type of the calibration tool may include an adhesion promoter facilitating contact between the surface of the support and the solid surface layer including the fluorescent material, which is in contact with the thin opaque mask A second type of the calibration tool may include the thin opaque mask fabricated (with or without an adhesion promoter) onto the support, and the solid surface layer including the fluorescent material located on the thin opaque mask

Claims

1. A calibration device for confirming or calibrating a biopolymeric array optical scanner, said device comprising:

a polymer layer comprising at least one fluorescent agent, wherein said device has minimal local and global nonuniformities and is dimensioned for placement in an optical scanner.

2. The device according to claim 1, wherein said at least one fluorescent agent is distributed substantially uniformly throughout said polymer.

3. The calibration device according to claim 1, wherein said polymer is a polyimide.

4. The calibration device according to claim 1, wherein the thickness of said polymer layer ranges from about 2 microns to about 250 microns.

5. The calibration device according to claim 1, wherein the thickness of said polymer layer ranges from about 1 micron to about 10 microns.

6. The calibration device according to claim 1, wherein said at least one fluorescent agent absorbs and emits light in the portion of the electromagnetic spectrum to which a photomultiplier tube of said optical scanner is sensitive.

7. The calibration device according to claim 1, wherein said at least one fluorescent agent absorbs and emits light in the wavelength range selected from the group consisting of ultraviolet, visible and near infrared.

8. The calibration device according to claim 1, wherein said polymer layer is a spin-coated polymer layer.

9. A method for calibrating a biopolymeric array optical scanning system, said method comprising:

(a) illuminating a surface of a calibration device with at least one light source, wherein said calibration device is a calibration device according to claim 1;

(b) obtaining fluorescence data from said surface of said calibration device; and

(c) calibrating said optical scanning system based upon said fluorescence data.

10. The method according to claim 9, wherein said step of illuminating comprises illuminating said surface of said calibration device in the portion of the electromagnetic spectrum to which a photomultiplier tube of said optical scanner is sensitive.

11. The method according to claim 9, wherein said step of illuminating comprises illuminating said surface of said calibration device in the wavelength range selected from the group consisting of ultraviolet, visible and near infrared.

12. The method according to claim 9, wherein said step of obtaining fluorescence data comprises detecting a signal related to the intensity of emitted light from said fluorescent agent.

13. The method according to claim 9, wherein said fluorescent agent(s) is distributed substantially uniformly throughout said surface.

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

(a) calibrating an optical scanner with a calibration device, wherein said calibration device is calibration device according to claim 1, (b) performing a hybridization assay with at least one array, and (c) scanning said array with said calibrated optical scanner.

15. A method for manufacturing a calibration device, said method comprising spin-coating a composition onto a substrate to produce a calibration device according to claim 1.

23. An optical scanner comprising a calibration device according to claim 1.

16. A calibration device for confirming or calibrating a biopolymeric array optical scanner, said device comprising:

a polymer layer comprising at least one fluorescent agent, wherein said device has minimal local and global nonuniformities; and

a transparent substrate;

wherein said device is dimensioned for placement in an optical scanner.

17. The calibration device according to claim 16, wherein said transparent substrate is glass.

18. The calibration device according to claim 17, wherein said at least one fluorescent agent is distributed substantially uniformly throughout said polymer.

19. The calibration device according to claim 16, wherein the thickness of said polymer layer ranges from about 1 micron to about 10 microns.

20. A calibration device for confirming or calibrating a biopolymeric array optical scanner, said device comprising:

a polymer layer comprising at least one fluorescent agent, wherein said device has minimal local and global nonuniformities; and

a substrate;

wherein said device is dimensioned for placement in an optical scanner.

21. The calibration device according to claim 20, wherein said at least one fluorescent agent is distributed substantially uniformly throughout said polymer.

22. The calibration device according to claim 20, wherein the thickness of said polymer layer ranges from about 1 micron to about 10 microns.

24. A calibration device for confirming or calibrating a biopolymeric array optical scanner, said device comprising:

a polymer layer comprising at least one fluorescent agent, wherein said device is dimensioned for placement in an optical scanner, and wherein said at least one fluorescent agent is distributed substantially uniformly throughout said polymer.

25. The device according to claim 24, wherein said polymer layer is of uniform thickness.