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

• US 9,638,637 B2
• Filed: 09/02/2014
• Issued: 05/02/2017
• 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): GUNBERG, EDWIN C
• Application Number:
  US14/474,742
• Publication Number:
  US 20140370612A1
• Time in Patent Office: 973 Days
• 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 for characterizing particles comprising;

moving a particle within a channel;

generating spatially modulated output light comprising moving the particle relative to a shadow mask disposed between the particle and a detector, the relative motion causing light emission from the particle to be spatially modulated;

recording a time modulated signal based on the spatially modulated output light, the time modulated signal including at least three transitions from an on state to an off state and including information about a position of the particle within the channel; and

detecting and evaluating the time modulated signal.

2. The method as set forth in claim 1 wherein the shadow mask is operative to modulate the signal obtained from the particle as a function of a position of a particle.

3. The method as set forth in claim 1 further comprising moving the shadow mask.

4. The method as set forth in claim 1 further comprising 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.

5. The method as set forth in claim 1 wherein the detecting comprises detecting with a pixilated detector.

6. 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 signal is used to determine DNA sequencing.

7. The method as set forth in claim 1 wherein the detecting comprises using a spectrometer to receive a fluorescent spectrum of the particle.

8. The method as set forth in claim 1 wherein the shadow mask is operative to pattern the light based on at least one of geometry, electric or magnetic field, fluorescence quenching, particle concentration, density, and acoustic standing wave.

9. The method as set forth in claim 1, wherein-generating the spatially modulated output light comprises generating a spatially modulated excitation region that includes an excitation pattern having at least three transitions from an area of relatively higher excitation to an area of relatively lower excitation.

10. The method as set forth in claim 9 wherein the generating of the excitation region comprises generating a spatially modulated pattern based on at least one of geometry, fluorescence quenching, analyte concentration, and density.

11. The method as set forth in claim 1 further comprising at least one of determining a location of the particle based on the time modulated signal, counting particles based on the time modulated signal, and sorting particles based on the time modulated signal.

12. The method as set forth in claim 1 wherein the detecting comprises detecting with a pixilated detector.

13. 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 signal is used to determine DNA sequencing.

14. The method as set forth in claim 1 wherein detecting comprises using a spectrometer to receive the fluorescent spectrum of the particle.

15. The method as set forth in claim 1 wherein detecting and evaluating the time modulated signal comprises detecting the time modulated signal in two-dimensions to locate the particle.

16. A method for characterizing particles comprising;

moving a particle within a channel;

generating spatially modulated output light comprising moving the particle relative to a shadow mask disposed between the particle and a detector, the relative motion causing light emission from the particle to be spatially modulated

recording a time modulated signal based on the spatially modulated output light, the time modulated signal including at least three transitions from an on state to an off state and including information about a position of the particle within the channel; and

detecting the time modulated signal.

17. A system for characterizing particles comprising;

a channel through which a particle can move;

a shadow mask configured to provide spatially modulated output light by spatially modulating light emission from the particle based on relative motion between the particle and the optical element; and

,

a detection system to record a time modulated signal based on the spatially modulated output light and evaluate the time modulated signal, the time modulated signal including at least three transitions from an on state to an off state and the time modulated signal including the at least three transitions including information about a position of the particle within the channel, wherein the shadow mask is disposed between the particle and the detection system.

18. The system as set forth in claim 17 further comprising an anti-resonant waveguide operative to cause the particle to emit light.

19. The system as set forth in claim 17 wherein the system is configured to determine a location of the particle based on the signal, count particles based on the signal, and sort particles based on the signal.

20. The system as set forth in claim 17 wherein the detection system allows for a two-dimensional evaluation.

21. The system as set forth in claim 17 wherein:

the shadow mask is operative to cause the particle to create time modulated signal comprises a spatially modulated excitation region that includes an excitation pattern having at least three transitions from an area of relatively higher excitation to an area of relatively lower excitation; and

the detection system is configured to record a time modulated signal generated by the particle as the particle moves through the excitation region and is excited upon exposure to the excitation region.

22. The system as set forth in claim 17 further comprising analyzer circuitry configured to perform at least one of determining a location of the particle based on the time modulated signal, counting particles based on the time modulated signal, and sorting particles based on the time modulated signal.