Method and system implementing spatially modulated excitation or emission for particle characterization with enhanced sensitivity

• US 8,821,799 B2
• Filed: 01/26/2007
• Issued: 09/02/2014
• Est. Priority Date: 01/26/2007
• Status: Active Grant

• Current Assignee: Xerox Corporation (Xerox Holdings Corp.)
• Original Assignee: Palo Alto Research Center, Inc. (Xerox Holdings Corp.)
• Inventors: Bassler, Michael, Kiesel, Peter, Schmidt, Oliver, Johnson, Noble M.
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US11/698,409
• Publication Number:
  US 20080181827A1
• Time in Patent Office: 2,776 Days
• Field of Search: None
• US Class Current: 422/105
• CPC Class Codes: G01N 15/1427 with the synchronisation of components, a time gate for operation of components, or suppression of particle coincidences
• CPC Class Codes: G01N 15/147 the analysis being performed on a sample stream
• CPC Class Codes: G01N 2015/1447 Spatial selection
• CPC Class Codes: G01N 2015/1454 using phase shift or interference, e.g. for improving contrast
• CPC Class Codes: G01N 21/6486 Measuring fluorescence of biological material, e.g. DNA, RNA, cells G01N21/6428 takes precedence
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]
• CPC Class Codes: Y10T 436/17 Nitrogen containing

Abstract

A method and system for using spatially modulated excitation/emission and relative movement between a particle (cell, molecule, aerosol, . . . ) and an excitation/emission pattern are provided. In at least one form, an interference pattern of the excitation light with submicron periodicity perpendicular to the particle flow is used. As the particle moves along the pattern, emission is modulated according to the speed of the particle and the periodicity of the stripe pattern. A single detector, which records the emission over a couple of stripes, can be used. The signal is recorded with a fast detector read-out in order to capture the “blinking” of the particles while they are moving through the excitation pattern. This concept enables light detection with high signal-to-noise ratio and high spatial resolution without the need of expensive and bulky optics.

Claims

1. A method of characterizing particles, the method comprising:

creating relative movement between at least one particle and an excitation region;

generating a spatially modulated excitation pattern within the excitation region, the spatially modulated excitation pattern including multiple high excitation areas interspersed with multiple low excitation areas, the spatially modulated excitation pattern extending along a length of the excitation region;

stimulating energy emission from the particle as the particle moves along the length of the excitation region;

detecting the energy emission from the particle using a detector;

generating a detector signal that is time modulated according to the spatially modulated excitation pattern;

analyzing the time modulated detector signal; and

characterizing the particle based on the analysis, wherein generating the spatially modulated excitation pattern within the excitation region comprises generating the spatially modulated excitation pattern based on at least one of an electric field, a magnetic field, and an acoustic standing wave.

2. The method as set forth in claim 1, wherein analyzing the time modulated detector signal comprises at least one of determining a location of the particle based on the signal, counting particles based on the signal, and sorting particles based on the signal.

3. The method as set forth in claim 1, wherein the detecting comprises detecting using a pixilated detector.

4. The method as set forth in claim 1,

wherein the particle is a portion of a DNA molecule or a molecule attached to the DNA molecule and the detector signal is used to determine a DNA sequence.

5. The method as set forth in claim 1, wherein the detecting comprises using a spectrometer to receive a fluorescent spectrum of a fluorescing analyte.

6. The method as set forth in claim 1, wherein generating the spatially modulated excitation pattern comprises generating a spatially modulated pattern based on environment.

7. The method as set forth in claim 1, wherein generating the spatially modulated excitation pattern extending along a length of the excitation region comprises:

generating a first spatially modulated excitation pattern extending along a length of the excitation region in a first direction; and

generating a second spatially modulated excitation pattern extending along a length of the excitation region in a second direction that is substantially orthogonal to the first direction.

8. The method as set forth in claim 1, wherein the energy emitted includes at least one of fluorescence excitation, elastic light scattering, inelastic light scattering, and chemo-luminescence.

9. The method of claim 1,

wherein analyzing the time modulated detector signal includes using a triangular fit function.

10. The method of claim 1, wherein analyzing the time modulated detector signal includes applying a fit function.

11. A method of characterizing particles, the method comprising:

generating a spatially modulated excitation pattern within an excitation region based on at least one of an electric field, a magnetic field, and an acoustic standing wave;

creating relative movement between at least one particle and the excitation region, the particle being excited upon exposure to the spatially modulated excitation pattern within the excitation region, the spatially modulated excitation pattern comprising multiple high excitation regions interspersed with multiple low excitation regions, the spatially modulated excitation pattern stimulating time modulated energy emission from the particle;

detecting the time modulated energy emission from the particle using a detector and generating a time modulated detector signal, the time modulated detector signal being time modulated based on the spatially modulated excitation pattern; and

extracting particle information by

detecting a presence of the particle by calculating an integral of the time modulated signal over a predetermined time period; and

triggering a positioning algorithm after the integral reaches a predetermined threshold, the positioning algorithm including applying a fit function to the time modulated signal.

12. A method comprising:

creating relative movement between at least one particle and an excitation region;

generating a spatially modulated excitation pattern within the excitation region based on a non-optical field, the spatially modulated excitation pattern including multiple high excitation areas interspersed with multiple low excitation areas, the spatially modulated excitation pattern extending along a length of the excitation region;

stimulating energy emission from the particle as the particle moves along the length of the excitation region;

detecting the energy emission from the particle using a detector; and

generating a detector signal that is time modulated according to the spatially modulated excitation pattern.

13. The method of claim 12, wherein the non-optical field is an electric field.

14. The method of claim 12, wherein the non-optical field is a magnetic field.

15. The method of claim 12, wherein the non-optical field is an acoustic standing wave.

16. The method of claim 12, wherein the particle is a portion of a DNA molecule or a molecule attached to the DNA molecule and the detector signal is used to determine a DNA sequence.

17. The method of claim 12, further comprising:

analyzing the time modulated detector signal; and

characterizing the particle based on the analysis.

18. The method of claim 17, wherein analyzing the time modulated detector signal comprises at least one of determining a location of the particle based on the signal, counting particles based on the signal, and sorting particles based on the signal.

19. The method of claim 17, wherein analyzing the time modulated detector signal includes applying a fit function.

20. The method of claim 19, wherein the fit function is a triangular fit function.

21. A method comprising:

creating relative movement between at least one particle and an excitation region;

generating a spatially modulated excitation pattern within the excitation region based on a non-optical field, the spatially modulated excitation pattern including multiple high excitation areas interspersed with multiple low excitation areas, the spatially modulated excitation pattern extending along a length of the excitation region; and

generating a detector signal that is time modulated according to the spatially modulated excitation pattern.

22. The method of claim 21, wherein the spatially modulated excitation pattern comprises electrical properties.

23. The method of claim 21, wherein the detector signal is based on capacitance.

24. The method of claim 21, wherein the detector signal is based on resistance.

25. The method of claim 21, wherein the detector signal is based on inductance.

26. The method of claim 21, wherein the detector signal is a current signal.

27. The method of claim 21, wherein the non-optical field is an electric field.

28. The method of claim 21, further comprising:

analyzing the time modulated detector signal including applying a fit function; and

characterizing the particle based on the analysis.