METHOD AND SYSTEM FOR STANDARIZING MICROSCOPE INSTRUMENTS

• US 20130310268A1
• Filed: 04/22/2013
• Published: 11/21/2013
• Est. Priority Date: 06/15/2007
• Status: Active Grant

• Current Assignee: Novartis Ag
• Original Assignee: HistoRx, Inc. (NeoGenomics, Inc.)
• Inventors: CHRISTIANSEN, Jason, PINARD, Robert, TEDESCHI, Gregory R., ZERKOWSKI, Meciej P.
• Granted Patent: US 9,080,978 B2
• Time in Patent Office: Days
• US Class Current: 506/9
• 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 standardizing a 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:

a. obtaining for the excitation light source, a light source correction factor;

b. applying the light source correction factor to the biological sample data, thereby obtaining a biological sample data standardized with regard to light intensity variability; and

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

wherein the light source correction factor comprises a correction factor for fluctuations in the intensity of the excitation light source.

2. The method of claim 1, in which the optical system is a microscope.

3. The method of claim 2, 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.

4. The method of claim 1, in which the biological sample data is obtained from an intensity measurement.

5. The method of claim 1, wherein obtaining the light source correction factor comprises collecting a relative light intensity measurement substantially coincidentally with the biological sample image.

6. The method of claim 1, in which the light source correction factor is obtained utilizing a calibration instrument installed in the optical system.

7. The method of claim 1 further comprising determining a calibration instrument compensation factor and applying the factor to the biological sample data.

8. The method of claim 1, in which the calibration instrument compensation factor is stored in the data storage portion of the optical system.

9. The method of claim 1 further comprising:

d. obtaining an optical system intrinsic factor; and

e. applying the optical system intrinsic factor to the biological sample data before determining the quantitative measurement of the standardized biological sample data, the biological sample data thereby standardized with regard to optical system intrinsic factors.

10. The method of claim 9, wherein obtaining the optical system intrinsic factor comprises:

d-1. illuminating by the excitation light source, a calibration biological sample configured to produce a standardized response;

d-2. obtaining in response to illuminating the calibration biological sample, measured biological sample data; and

d-3. determining the optical system intrinsic factor indicative of relative performance of the optics portion.

16. The method of claim 1, in which 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, or combinations of one or more of any of the foregoing.

19. The method of claim 1, in which applying the light source correction factor to the biological sample data comprises applying the light source correction factor to one or more individual channels of the biological sample data.

11. 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, a plurality of channels and a data storage portion, cooperatively arranged for obtaining biological sample data, comprising:

a. obtaining an optical system intrinsic factor for each individual channel;

b. applying the optical system intrinsic factor for a particular channel to the biological sample data from the same channel, thereby obtaining a biological sample data measurement standardized with regards to optical system intrinsic factors; and

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

12. The method of claim 11 further comprising:

d. obtaining a light source correction factor; and

e. applying the correction factor to the 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.

17. The method of claim 11, in which 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, or combinations of one or more of any of the foregoing.

13. A computer-usable medium having computer readable instructions stored thereon for execution by a processor to perform a method for standardizing a 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, where the instructions comprise the steps of:

a. obtaining for the excitation light source, a light source correction factor;

b. applying the light source correction factor to the biological sample data, thereby obtaining a biological sample data standardized with regard to light intensity variability; and

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

wherein the light source correction factor comprises a correction factor for fluctuations in the intensity of the excitation light source.

18. The method of claim 13, in which 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, or combinations of one or more of any of the foregoing.

14. 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 excitation light source, a light source correction factor;

means for applying the light source correction factor to the biological sample data, thereby obtaining a biological sample data standardized with regard to light intensity variability; and

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

wherein the light source correction factor comprises a correction factor for fluctuations in the intensity of the excitation light source.

15. A method for standardizing a 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:

a. obtaining for the excitation light source, a light source correction factor;

b. applying the light source correction factor to the biological sample data, thereby obtaining a biological sample data standardized with regard to light intensity variability; and

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

wherein obtaining the light source correction factor comprises collecting a relative light intensity measurement substantially simultaneously with the biological sample image.

20. The method of claim 15, in which 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, or combinations of one or more of any of the foregoing.