Method and system for standardizing microscope instruments

• US 9,080,978 B2
• Filed: 04/22/2013
• Issued: 07/14/2015
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Novartis Ag
• Original Assignee: Novartis Ag
• Inventors: Pinard, Robert, Zerkowski, Maciej P., Tedeschi, Gregory R., Christiansen, Jason
• Primary Examiner(s): Geisel, Kara E
• Assistant Examiner(s): Underwood, Jarreas C
• Application Number:
  US13/867,817
• Publication Number:
  US 20130310268A1
• Time in Patent Office: 813 Days
• Field of Search: 3562371-2416, 3562421-2438, 356426-431, 356600-640
• US Class Current: 1/1
• CPC Class Codes: G01N 2021/6439 with indicators, stains, dyes, tags, labels, marks
• CPC Class Codes: G01N 21/276 with alternation of sample and standard in optical path
• CPC Class Codes: G01N 21/278 Constitution of standards
• CPC Class Codes: G01N 21/6458 Fluorescence microscopy fluorescence microscopes per se G02B21/0076 and G02B21/16
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence

Abstract

Methods and apparatus for standardizing quantitative measurements from a microscope system. The process includes a calibration procedure whereby an image of a calibration slide is obtained through the optics of the microscope system. The calibration slide produces a standard response, which can be used to determine a machine intrinsic factor for the particular system. The machine intrinsic factor can be stored for later reference. In use, images are acquired of a target sample and of the excitation light source. The excitation light source sample is obtained using a calibration instrument configured to sample intensity. The calibration instrument has an associated correction factor to compensate its performance to a universally standardized calibration instrument. The machine intrinsic factor, sampled intensity, and calibration instrument correction factor are usable to compensate a quantitative measurement of the target sample in order to normalize the results for comparison with other microscope systems.

Claims

1. A method for obtaining a quantitative standardized biological sample data measurement from an optical system having an excitation light source, an optics portion, a detection portion, and a data storage portion, cooperatively arranged for obtaining biological sample data, comprising:

a. obtaining an optical system intrinsic factor, based on a calibration surface configured to produce a standardized response;

b. applying the optical system intrinsic factor to a biological sample data, thereby obtaining a biological sample data measurement standardized with regards to optical system variability; and

c. determining a quantitative measure of the standardized biological sample data.

2. The method of claim 1 further comprising:

d. obtaining a light source correction factor; and

e. applying the correction factor to a biological sample measurement standardized with regard to optical system intrinsic factors, thereby obtaining a biological sample data measurement further standardized with regard to light intensity variability.

3. The method of claim 1 further comprising obtaining for the excitation light source, a light source correction factor.

4. The method of claim 3, wherein the light source correction factor is obtained by positioning the calibration surface along an optical path.

5. The method of claim 1, wherein the optical system further comprises a plurality of channels, and the optical system intrinsic factor is obtained for a particular channel.

6. The method of claim 1, wherein the image capture portion is a digital camera.

7. The method of claim 1, wherein the optical system is a microscope.

8. The method of claim 7, wherein the microscope is selected from the group consisting of:

optical microscopes using incoherent illumination sources;

optical microscopes using polarized illumination sources;

optical microscopes using fluorescent illumination sources;

microscopes using coherent illumination sources; and

combinations thereof.

9. The method of claim 1, wherein the biological sample is selected from the group consisting of a cell, a group of cells, a tissue section, an array of tissue sections, a micro tissue array, and combinations of one or more of any of the foregoing.

10. The method of claim 1, wherein the biological sample data is obtained from an intensity measurement.

18. The method of claim 1, wherein the biological sample data is an image of a biological sample.

11. A microscope system for obtaining a standardized quantitative measurement, of biological sample data imaged by an optical system having an excitation light source, an optics portion, an image capture portion and a data storage portion, cooperatively arranged for obtaining an image of the biological sample, comprising:

means for obtaining for the optical system, an optical system intrinsic factor, based on a calibration surface configured to produce a standardized response;

means for applying the optical system intrinsic factor to a biological sample data, thereby obtaining a biological sample data measurement standardized with regards to optical system variability; and

means for determining a quantitative measure of the standardized biological sample data.

12. The microscope system of claim 11 further comprising means for obtaining for the excitation light source, a light source correction factor.

13. The microscope system of claim 12 wherein obtaining the means for the obtaining the excitation light source includes means for positioning the calibration surface along an optical path.

14. The microscope system of claim 11, wherein the optical system further comprises a plurality of channels, and the optical system intrinsic factor is obtained for a particular channel.

15. The microscope system of claim 11, wherein the image capture portion is a digital camera.

16. The microscope system of claim 11, wherein the biological sample is selected from the group consisting of a cell, a group of cells, a tissue section, an array of tissue sections, a micro tissue array, and combinations of one or more of any of the foregoing.

17. The microscope system of claim 11, wherein the biological sample data is obtained from an intensity measurement.