System and method for classifying cells and the pharmaceutical treatment of such cells using Raman spectroscopy

• US 20070153268A1
• Filed: 01/05/2007
• Published: 07/05/2007
• Est. Priority Date: 01/05/2006
• Status: Active Grant

• Current Assignee: Chemimage Corporation
• Original Assignee: Chemimage Corporation
• Inventors: Maier, John, Neiss, Jason, Panza, Janice
• Granted Patent: US 7,570,356 B2
• Time in Patent Office: Days
• US Class Current: 356/301
• CPC Class Codes: G01J 3/28 Investigating the spectrum using colour filters G01J3/51
• CPC Class Codes: G01J 3/36 Investigating two or more bands of a spectrum by separate detectors
• CPC Class Codes: G01J 3/44 Raman spectrometry; Scattering spectrometry ; Fluorescence spectrometry
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1488 Methods for deciding
• CPC Class Codes: G01N 2021/656 Raman microprobe
• CPC Class Codes: G01N 21/65 Raman scattering

Abstract

A system and method to distinguish normal cells from apoptotic cells. A pre-determined vector space is selected where the vector space mathematically describes a first plurality of reference Raman data sets for normal cells and a second plurality of reference Raman data sets for apoptotic cells. A sample is irradiated with substantially monochromatic light generating a target Raman data set based on scattered photons. The target Raman data set is transformed into a vector space defined by the pre-determined vector space. A distribution of transformed data is analyzed in the pre-determined vector space. Based on the analysis, the sample is classified as containing normal cells, apoptotic cells, and a combination of normal and apoptotic cells. The sample includes the step of treating the sample with a pharmaceutical agent prior to irradiating the sample. Based on the classification, the therapeutic efficiency of the pharmaceutical agent is assessed.

Claims

1. A method comprising:

selecting a pre-determined vector space that mathematically describes a first plurality of reference Raman data sets for normal cells and a second plurality of reference Raman data sets for apoptotic cells;

irradiating a sample containing at least one cell with substantially monochromatic light to thereby generate scattered photons;

collecting a target Raman data set based on said scattered photons;

transforming the target Raman data set into said pre-determined vector space;

analyzing a distribution of transformed data in the pre-determined vector space; and

based on said analysis, classifying the at least one cell as a normal cell or an apoptotic cell.

2. The method of claim 1, wherein said analyzing is performed by calculating a Mahalanobis distance between the target Raman data set transformed into said vector space and one or more of the following:

the first plurality of reference Raman data sets in said pre-determined vector space; and

the second plurality of reference Raman data sets in said predetermined vector space.

3. The method of claim 1, wherein said first reference set of Raman data sets corresponds to one or more of the following:

a plurality of first reference Raman spectra, wherein each spectrum corresponding to a reference normal cell; and

a plurality of first reference spatially accurate wavelength resolved Raman images, wherein each image corresponding to a reference normal cell.

4. The method of claim 1, wherein said second reference set of Raman data sets corresponds to one or more of the following:

a plurality of second reference Raman spectra, wherein each spectrum corresponding to a reference apoptotic cell; and

a plurality of second reference spatially accurate wavelength resolved Raman images, wherein each image corresponding to a reference apoptotic cell.

5. The method of claim 1, wherein said target Raman data set corresponds to one or more of the following:

a plurality Raman spectra of the sample; and

a plurality of spatially accurate wavelength resolved Raman images of the sample.

6. The method of claim 1, further comprising:

treating said sample with a pharmaceutical agent prior to irradiating said sample; and

based on said classifying, assessing a therapeutic efficiency of said pharmaceutical agent.

7. The method of claim 6, wherein said treatment results in cell apoptosis.

8. The method of claim 6, wherein said treatment prevents cell apoptosis.

9. The method of claim 1, further comprising:

labeling said sample with a fluorescent material;

obtaining a fluorescence emission signature of said fluorescent labeled sample and

determining whether said at least one cell expresses phosphatidylserine and a location of said at least one cell in said sample based on said fluorescence signature of the sample.

10. The method of claim 9, wherein said collecting further comprising:

collecting said target Raman data set based on said scattered photons generated by said location.

11. A method comprising:

selecting a pre-determined vector space that mathematically describes a first plurality of reference Raman data sets for normal cells and

a second plurality of reference Raman data sets for apoptotic cells;

irradiating a biological sample with substantially monochromatic light to thereby generate scattered photons;

collecting a target Raman data set based on said scattered photons;

transforming the target Raman data set into a vector space defined by the pre-determined vector space;

analyzing a distribution of transformed data in the pre-determined vector space; and

based on said analyzing, classifying the biological sample as comprised of one of the following;

normal cells, apoptotic cells, and a combination of normal and apoptotic cells.

12. The method of claim 11, further comprising:

treating said sample with a pharmaceutical agent prior to irradiating said sample; and

based on said classifying, assessing a therapeutic efficiency of said pharmaceutical agent.

13. The method of claim 11, wherein said target Raman data set corresponds to one or more of the following:

a plurality Raman spectra of the sample; and

a plurality of spatially accurate wavelength resolved Raman images of the sample.

14. The method of claim 11, further comprising:

obtaining a fluorescence emission signature of a fluorescent labeled biological sample; and

determining whether said sample expresses phosphatidylserine and a location at least one cell in said sample based on said fluorescence signature of said biological sample.

15. The method of claim 11, wherein said analyzing is performed by calculating a Mahalanobis distance between the target Raman data set transformed into said vector space and one or more of the following:

the first plurality of reference Raman data sets in said pre-determined vector space.

16. A system comprising:

a first database having a first plurality of reference Raman data sets for normal cells;

a second database having a second plurality of reference Raman data sets for apoptotic cells;

an illumination source;

a spectroscopic device;

a machine readable program code containing executable program instructions; and

a processor operatively coupled to the illumination source and the spectroscopic device, and configured to execute said machine readable program code so as to perform the following;

select a pre-determined vector space that mathematically describes the first plurality of reference Raman data sets for normal cells and the second plurality of reference Raman data sets for apoptotic cells;

configure said illumination source to illuminate a biological sample with substantially monochromatic light to thereby generate scattered photons;

configure said spectroscopic device to collect a Raman data set based on said scattered photons;

transform the target Raman data set into a vector space defined by the pre-determined vector space;

analyze a distribution of transformed data in the pre-determined vector space;

based on said analysis, classify the biological sample as comprised of one of the following;

normal cells, apoptotic cells, and a combination of normal and apoptotic cells.

17. The system of claim 16, wherein said biological sample is treated with a pharmaceutical agent prior to illumination thereof by said illumination source, and wherein

said processor is further configured to perform the following:

based on said classification, assess a therapeutic efficiency of said pharmaceutical agent.

18. A system comprising:

a first database having a first plurality of reference Raman data sets for normal cells;

a second database having a second plurality of reference Raman data sets for apoptotic cells;

means to irradiate a biological sample with substantially monochromatic light to thereby generate scattered photons;

means to collect a target Raman data set based on said scattered photons;

means to transform the target Raman data set into a vector space defined by the pre-determined vector space;

means to analyze a distribution of transformed data in the pre-determined vector space; and

based on said analysis, means to classify the biological sample as comprised of normal cells or apoptotic cells.

19. A storage medium containing machine readable program code, which, when executed by a processor, causes said processor to perform the following:

select a pre-determined vector space that mathematically describes a first plurality of reference Raman data sets for normal cells and a second plurality of reference Raman data sets for apoptotic cells;

configure an irradiation source to irradiate a biological sample cell with substantially monochromatic light to thereby generate scattered photons;

configure a spectroscopic device to collect a target Raman data set based on said scattered photons;

transform the target Raman data set into a vector space defined by the pre-determined vector space;

analyze a distribution of transformed data in the pre-determined vector space; and

based on said analysis, classify the biological sample as comprised of normal cells or apoptotic cells.