Method and system for molecular array scanner calibration

• US 6,929,951 B2
• Filed: 02/28/2002
• Issued: 08/16/2005
• Est. Priority Date: 02/28/2002
• Status: Expired due to Fees

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Corson, John F., Parker, Russell A., Dorsel, Andreas N., Chow, Andre B.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US10/086,932
• Publication Number:
  US 20030165871A1
• Time in Patent Office: 1,265 Days
• Field of Search: 436/8, 436/164, 436/172, 702/19, 702/22, 702/27, 702/28, 702/32, 422/82.05, 422/82.08, 422/82.09, 73/10.1, 435/6, 435/287.2, 435/7.9
• US Class Current: 436/8
• 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 21/6428 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: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing

Abstract

A method and system for calibrating molecular arrays to a reference molecular array, and for subsequently calibrating the molecular arrays to maintain a constant signal-intensity-to-label-concentration ratio. In the first step of the two-step calibration method, a reference array coated with the fluorophore or chromophore used to label probe molecules is employed, while in the second step of the two-step method, a reference array coated with a stable dye is employed.

Claims

1. A method for calibrating a first molecular array scanner with a second, reference molecular array scanner, the method comprising:

initially using a same-dye reference array comprising a first dye to calibrate the first molecular array scanner with the second, reference molecular array scanner; and

maintaining the initial calibration of said first molecular array scanner with said reference molecular array scanner by scanning a stable-dye reference array comprising a second dye with the first molecular array scanner.

2. The method of claim 1 wherein said same-dye reference array is used to initially calibrate the first molecular array scanner with the second, reference molecular array scanner by:

scanning the same-dye reference array in the second, reference molecular array scanner to determine a measured signal intensity for the same-dye reference array in the reference molecular array scanner;

calculating an expected intensity for scanning the same-dye reference array in the second, reference molecular array scanner a second time; and

scanning the same-dye reference array in the first scanner, and adjusting parameters in the first molecular array scanner to produce the expected intensity.

3. The method of claim 2 wherein calculating an expected intensity for scanning the same-dye reference array in the second, reference molecular array scanner a second time further comprises:

determining a function of expected intensity per scan of the same-dye reference array; and

selecting the expected intensity corresponding to one more than a number of times that the same-dye reference array has been scanned.

4. The method of claim 1 wherein maintaining the initial calibration further includes:

following initial calibration, scanning a stable-dye reference array with the first molecular array scanner in order to determine a signal-intensity-to-stable-dye-concentration ratio; and

periodically rescanning the stable-dye reference array with the first molecular array scanner, and adjusting the first molecular array scanner to provide the determined signal-intensity-to-stable-dye-concentration ratio.

5. A system for calibrating a number of molecular array scanners to provide a fixed signal-intensity-to-label-concentration ratio, the system comprising:

a reference molecular array scanner;

a number of molecular array scanners to be calibrated by the reference molecule array scanner

a same-dye reference array comprising a first dye used to establish an initial calibration of each of the number of molecular array scanners to the reference molecular array scanner; and

a stable-dye reference array comprising a second dye used to maintain the initiated calibration.

6. The system of claim 5 configured so that the same-dye reference array is used to establish an initial calibration of a first molecular array scanner to the reference molecular array scanner by:

scanning the same-dye reference array in the reference molecular array scanner to determine a measured signal intensity for the same-dye reference array in the reference molecular array scanner;

calculating an expected intensity for scanning the same-dye reference array in the reference molecular array scanner a second time; and

adjusting the first molecular array scanner to produce the respective calculated expected intensity.

7. The system of claim 6 wherein calculating the expected intensity for scanning the same-dye reference array in the reference molecular array scanner a second time further comprises:

determining a function of expected intensity per scan of the same-dye reference array; and

selecting the expected intensity for the first molecular array scanner corresponding to one more than a number of times that the same-dye reference array has been scanned.

8. The system of claim 5 configured so that one or more stable-dye reference arrays are used to maintain the initial calibration of the first molecular array scanner by:

initial calibration, scanning a stable-dye reference array with the first molecular array scanner in order to determine a signal-intensity-to-stable-dye-concentration ratio; and

periodically rescanning the stable-dye reference array with the first molecular array scanner, and adjusting the first molecular array scanner to provide the determined signal-intensity-to-stable-dye-concentration ratio.