Modifying the output of a laser to achieve a flat top in the laser's Gaussian beam intensity profile

• US 8,451,524 B2
• Filed: 09/20/2010
• Issued: 05/28/2013
• Est. Priority Date: 09/29/2009
• Status: Active Grant

• Current Assignee: Cytek Biosciences Inc
• Original Assignee: Amnis Corporation (DiaSorin S.p.A.)
• Inventors: Ortyn, William, Perry, David, Montague, Tom
• Primary Examiner(s): FONT, FRANK G
• Application Number:
  US12/886,182
• Publication Number:
  US 20110085221A1
• Time in Patent Office: 981 Days
• Field of Search: 359/224.1, 359/224.2, 359/199.1, 372/29.01, 372/29.014, 372/31, 372/32, 356/28, 356/28.5, 356/39, 356/73, 356/301, 356/317, 356/326, 356/338, 356/400, 356/419, 356/445, 2502012-2014
• US Class Current: 359/224.1
• CPC Class Codes: G01N 15/1434 Optical arrangements
• CPC Class Codes: G01N 15/1459 the analysis being performed on a sample stream
• CPC Class Codes: G02B 19/0014 at least one surface having optical power
• CPC Class Codes: G02B 19/0052 the light source comprising a laser diode coupling into light guides using intermediate optical elements G02B6/4204; semiconductor lasers having optical devices external to the laser cavity H01S5/005
• CPC Class Codes: G02B 26/0858 the reflecting means being moved or deformed by piezoelectric means
• CPC Class Codes: G02B 26/105 with one or more pivoting mirrors or galvano-mirrors G02B26/101 takes precedence
• CPC Class Codes: G02B 27/0927 Systems for changing the beam intensity distribution, e.g. Gaussian to top-hat
• CPC Class Codes: G02B 27/0944 Diffractive optical elements, e.g. gratings, holograms gratings per se G02B5/18; holograms used as optical elements per se G02B5/32
• CPC Class Codes: G02B 27/0977 Reflective elements
• CPC Class Codes: G02F 1/11 based on acousto-optical elements, e.g. using variable diffraction by sound or like mechanical waves acousto-optical deflection G02F1/33
• CPC Class Codes: G02F 2203/18 adaptive optics, e.g. wavefront correction

Abstract

A laser beam is periodically deflected before being directed into a sample volume. The beam is deflected at a frequency such that the beam makes one or more passes through the sample volume while data are collected from the sample volume. The amplitude of motion of the beam, the dwell time of the beam at any given point, and the Gaussian intensity profile of the beam cooperate to produce an effective flat topped illumination profile for the light that is incident on specimens in the sample volume. The total photon exposure at any given point in the sample volume is a function of both the beam intensity and the dwell time at that location. Therefore, a longer dwell time and lower intensity at the edge of the profile are in balance with a shorter dwell time and higher intensity at the center of the profile.

Claims

1. Apparatus for generating an effective flat top intensity profile for light illuminating moving particles in a particle analyzer, comprising:

(a) a source of light;

(b) at least one optical component for focusing the light from the source as a beam that is incident on a particle moving through the particle analyzer; and

(c) a scanner for scanning the light across the particle at a scan frequency selected so that the particle is illuminated with at least one cycle of the light being scanned, and with an amplitude for the scanning that is selected to be less than a diameter of the beam where it is incident on the particle and so that the light remains incident on the particle as the particle passes through a portion of the particle analyzer, wherein the scan frequency, the amplitude for the scanning, and an intensity profile of the source of light cooperate to generate an effective flat top intensity profile for the light being scanned.

2. The apparatus of claim 1, wherein the source of light comprises a laser.

3. The apparatus of claim 1, wherein the scanner comprises an item selected from the group consisting of:

(a) an acousto-optical modulator;

(b) an acousto-optical deflector;

(c) a reflective device that is moved periodically; and

(d) a diffractive device.

4. The apparatus of claim 1, wherein the scanner includes a reflective surface and an actuator for moving the reflective surface to scan the light reflected from the reflective surface.

5. The apparatus of claim 4, wherein the reflective surface comprises a mirror that is supported by at least one flexure that enables the mirror to be deflected in a periodic motion, when driven by the actuator.

6. The apparatus of claim 4, wherein the actuator comprises at least one piezoelectric transducer.

7. The apparatus of claim 1, wherein the scanner is driven to deflect the light being scanned at about a resonant frequency of the scanner.

8. The apparatus of claim 1, wherein the at least one optical component comprises one or more lenses that focus a beam of the light from the scanner onto the particle in the particle analyzer, reducing a diameter of beam and increasing an intensity of the light incident on the particle.

9. The apparatus of claim 1, wherein a rate at which the scanner scans the light changes as a function of a frequency of a drive signal that is applied to the scanner to drive it.

10. The apparatus of claim 1, wherein a motion profile of the scanned beam varies in according with at least one motion profile selected from the group consisting of:

(a) sinusoidal motion profile;

(b) a linear motion profile;

(c) a saw tooth motion profile; and

(d) a modified sinusoidal motion profile.

11. The apparatus of claim 1, wherein the scanner includes a reflective surface that is driven to move so that the amplitude of the scanning where the beam is incident on the particle is in the range from about 4.5 microns to about 12.0 microns.

12. The apparatus of claim 1, wherein the scanner directs the beam of the light that is scanned onto a portion of the particle analyzer through which a plurality of particles pass in succession, so that the plurality of particles are scanned by the beam.

13. A method for generating a flat top intensity profile for light illuminating moving particles in a particle analyzer, comprising the steps of:

(a) directing light from a source toward a scanner;

(b) scanning the light from the source that is incident on the scanner, to produce scanned light, the scanner being driven to scan a beam of the light at a desired frequency and with a desired amplitude; and

(c) focusing the scanned light onto a portion of the particle analyzer through which a particle is disposed, such that a particle in the portion of the particle analyzer is illuminated, wherein the desired frequency is selected so that the particle is illuminated with at least one cycle of the scanned light, and the desired amplitude for the scanning is selected to be less than a cross-sectional dimension of the beam, both the desired amplitude and the cross-sectional dimension of the beam being measured at a point where the beam is incident on the particle, and so that the beam remains incident on the particle as the particle is scanned by the beam within the portion of the particle analyzer, the desired frequency, the desired amplitude for the scanning, and an intensity profile of the source of the light cooperating to generate a flat top intensity profile for the scanned light that is focused on the particle.

14. The method of claim 13, further comprising the step of producing the light using a laser source.

15. The method of claim 13, wherein the step of scanning comprises the step of deflecting the beam using either an acousto-optical modulator, an acousto-optical deflector, a moving reflective surface, or a diffractive device.

16. The method of claim 13, wherein the step of scanning comprises the step of scanning the light from a reflective surface that is moved by an actuator in accord with a desired motion profile.

17. The method of claim 16, wherein the step of scanning comprises the step of supporting a mirror comprising the reflective surface using at least one flexure that enables the mirror to be deflected when moved by the actuator.

18. The method of claim 16, wherein the step of scanning comprises the step of applying a drive signal to an actuator comprising at least one piezoelectric transducer.

19. The method of claim 13, wherein the step of scanning comprises the step of scanning the light at about a resonant frequency of the scanner.

20. The method of claim 13, wherein the step of focusing the scanned light comprises the step of positioning at least one optical component between the scanner and the portion of the particle analyzer through which the particle is passing, the at least one optical component comprising one or more lenses that focus a beam of the light from the scanner onto the particle as the particle is passing through the particle analyzer, reducing a diameter of the beam and increasing an intensity of the light that is incident on the particle.

21. The method of claim 13, wherein the step of scanning comprises the step of changing a rate at which the scanner scans the light as a function of a drive signal that is applied to the scanner to drive it.

22. The method of claim 13, wherein the step of scanning comprises the step of driving the scanner to scan the light with at least one motion profile selected from the group consisting of:

(a) sinusoidal motion profile;

(b) a linear motion profile;

(c) a saw tooth motion profile; and

(d) a modified sinusoidal motion profile.

23. The method of claim 13, wherein the step of scanning comprises the step of driving a reflective surface that reflects the light to move with an amplitude that is in the range from about 4.5 microns to about 12.0 microns.

24. The method of claim 13, wherein the step of scanning comprises the step of directing the scanned light onto a portion of the particle analyzer through which a plurality of particles are moving.

25. Apparatus for producing light having an effective flat top intensity profile for illuminating a sample disposed within a particle analyzer, comprising:

(a) a source of light;

(b) a deflecting component that is mounted to receive incident light from the source and to deflect the incident light in a periodic manner, to produce a beam of scanned light that is directed toward a sample volume of the particle analyzer, the deflecting component being driven to scan the beam at a frequency selected so as to illuminate a sample in the sample volume of the particle analyzer with at least one cycle of the scanned light and with an amplitude selected to be less than a cross-sectional dimension of the beam of the scanned light as measured at a point where the beam is incident on the sample and so that the sample is fully illuminated with the scanned light while the sample is in the sample volume of the particle analyzer; and

(c) one or more lenses that focus the scanned light onto a sample in the sample volume of the particle analyzer, a peak of an intensity profile of the light produced by the source being of a lower magnitude than a peak of the effective flat topped intensity profile of the scanned light that illuminates the sample volume, due to an increased intensity of the scanned light caused by focusing the scanned light with the one or more lenses and the flat topped intensity profile of the scanned and focused light being caused by scanning the light with the deflecting component at the frequency and amplitude.