Illumination apparatus optimized for synthetic aperture optics imaging using minimum selective excitation patterns

• US 9,772,505 B2
• Filed: 03/02/2016
• Issued: 09/26/2017
• Est. Priority Date: 03/19/2010
• Status: Active Grant

• Current Assignee: Rebus Biosystems, Inc.
• Original Assignee: Optical Biosystems Incorporated
• Inventors: Lee, Chun-Sheu, Park, Jong Buhm
• Primary Examiner(s): ELAHI, SHAN E
• Application Number:
  US15/059,245
• Publication Number:
  US 20160178921A1
• Time in Patent Office: 573 Days
• Field of Search: 348 80
• CPC Class Codes: G02B 21/06 Means for illuminating specimens
• CPC Class Codes: G02B 21/18 Arrangements with more than one light path, e.g. for comparing two specimens
• CPC Class Codes: G02B 21/367 providing an output produced by processing a plurality of individual source images, e.g. image tiling, montage, composite images, depth sectioning, image comparison
• CPC Class Codes: G02B 27/58 Optics for apodization or superresolution; Optical synthetic aperture systems
• CPC Class Codes: G06T 2207/10056 Microscopic image
• CPC Class Codes: G06T 2207/30072 Microarray; Biochip, DNA array; Well plate
• CPC Class Codes: G06T 5/00 Image enhancement or restoration
• CPC Class Codes: G06T 5/50 using two or more images, e.g. averaging or subtraction
• CPC Class Codes: G06T 7/0012 Biomedical image inspection
• CPC Class Codes: G06V 20/693 Acquisition
• CPC Class Codes: G06V 20/695 Preprocessing, e.g. image segmentation

Abstract

A synthetic aperture optics (SAO) imaging method minimizes the number of selective excitation patterns used to illuminate the imaging target, based on the objects'"'"' physical characteristics corresponding to spatial frequency content from the illuminated target and/or one or more parameters of the optical imaging system used for SAO. With the minimized number of selective excitation patterns, the time required to perform SAO is reduced dramatically, thereby allowing SAO to be used with DNA sequencing applications that require massive parallelization for cost reduction and high throughput. In addition, an SAO apparatus optimized to perform the SAO method is provided. The SAO apparatus includes a plurality of interference pattern generation modules that can be arranged in a half-ring shape.

Claims

1. A method for performing synthetic aperture optics (SAO) on a target including one or more objects, the method comprising the steps of:

illuminating the target with a predetermined number (N) of selective excitation patterns, the N selective excitation patterns determined based on a minimum distance (Δ

x) between the objects to be resolved by SAO and a width (w) of a detected area captured by a pixel of an optical imaging system for optical imaging of the illuminated target;

optically imaging the illuminated target at a resolution insufficient to resolve the objects, the illuminated target being imaged by the optical imaging system through a lens included in an imaging path that is physically separated from illumination paths through which the N selective excitation patterns are delivered to the target; and

processing optical images of the illuminated target using information on the selective excitation patterns to obtain a final image of the illuminated target at a resolution sufficient to resolve the objects.

2. The method of claim 1, wherein the number (N) of selective excitation patterns corresponds to the number of k-space sampling points in a k-space sampling space in a frequency domain, an extent of the k-space sampling space being substantially proportional to an inverse of the minimum distance (Δ

x) between the objects that is to be resolved by SAO, and an inverse of a k-space sampling interval between the k-space sampling points being less than the width (w) of a detected area captured by the pixel of the optical imaging system.

3. The method of claim 2, wherein the number (N) of selective excitation patterns is determined from:

N=floor(L/2)

wherein L is the number of the k-space sample points in the k-space sampling space, and floor ( ) rounds the number to the nearest but smaller integer; and

L=round((PFOV/Δ

x)²)

wherein round ( ) rounds the number to the nearest integer, and PFOV is the extent in a reciprocal space of the k-space to be reconstructed from samples in the k-space sampling space.

4. The method of claim 2, further comprising removing aliasing in the final image caused as a result of using said inverse of the k-space sampling interval less than the width (w) of the detected area captured by the pixel of said optical imaging system.

5. The method of claim 1, further comprising removing effects of apodization caused by a weighting function of the optical imaging system, said weighting function corresponding to a convolution of a pixel sensitivity function of the pixel of the optical imaging system for detecting light from the illuminated target and a point-spread function of the lens through which said light is passed through in the optical imaging system.

6. The method of claim 1, wherein the number (N) of selective excitation patterns is determined based upon at least one of a plurality of parameters of the optical imaging system, including the width (w) of the detected area captured by the pixel of the optical imaging system, a wavelength of light from the illuminated target, and a numerical aperture of the lens through which said light is passed through in the optical imaging system.

7. The method of claim 1, wherein the number (N) of selective excitation patterns corresponds to non-uniformly selected ones of the k-space sampling points.

8. The method of claim 1, wherein the target is illuminated with two phases of each one of the selective excitation patterns and another phase of only one of the selective excitation patterns, said another phase being different from the two phases.

9. The method of claim 1, wherein the target is illuminated with three phases of each one of the selective excitation patterns.

10. A system for performing synthetic aperture optics (SAO) on a target including one or more objects, the system comprising:

a plurality of illumination devices configured to illuminate the target with a predetermined number (N) of selective excitation patterns, the N selective excitation patterns determined based on a minimum distance (Δ

x) between the objects to be resolved by SAO and a width (w) of a detected area captured by a pixel of an optical imaging device for optical imaging of the illuminated target;

the optical imaging device configured to optically image the illuminated target at a resolution insufficient to resolve the objects, the illuminated target being imaged by the optical imaging device through a lens included in an imaging path that is physically separated from illumination paths through which the N selective excitation patterns are delivered to the target; and

a SAO processing device configured to process the optical images of the illuminated target using information on the selective excitation patterns to obtain a final image of the illuminated target at a resolution sufficient to resolve the objects.

11. The system of claim 10, wherein the number (N) of selective excitation patterns corresponds to the number of k-space sampling points in a k-space sampling space in a frequency domain, an extent of the k-space sampling space being substantially proportional to an inverse of the minimum distance (Δ

x) between the objects that is to be resolved by SAO and an inverse of a k-space sampling interval between the k-space sampling points being less than the width (w) of the detected area captured by the pixel of the optical imaging device.

12. The system of claim 11, wherein the number (N) of selective excitation patterns is determined from:

N=floor(L/2)

wherein L is the number of the k-space sample points in the k-space sampling space, and floor ( ) rounds the number to the nearest but smaller integer; and

L=round((PFOV/Δ

x)²)

wherein round ( ) rounds the number to the nearest integer, and PFOV is the extent in a reciprocal space of the k-space to be reconstructed from samples in the k-space sampling space.

13. The system of claim 11, wherein the SAO processing device is further configured to remove aliasing in the final image caused as a result of using said inverse of the k-space sampling interval less than the width (w) of the detected area captured by the pixel of the optical imaging device.

14. The system of claim 10, wherein the SAO processing device is further configured to remove effects of apodization caused by a weighting function of the optical imaging device, said weighting function corresponding to a convolution of a pixel sensitivity function of a pixel of said optical imaging device for detecting light from the illuminated target and a point-spread function of the lens through which said light is passed through in said optical imaging device.

15. The system of claim 10, wherein the number (N) of selective excitation patterns is determined based upon at least one of a plurality of parameters of said optical imaging device, including the width (w) of the detected area captured by the pixel of said optical imaging device, a wavelength of the light from the illuminated target, and a numerical aperture of the lens through which said light is passed through in said optical imaging device.

16. The system of claim 10, wherein the number (N) of selective excitation patterns corresponds to non-uniformly selected ones of the k-space sampling points.

17. The system of claim 10, wherein the target is illuminated with two phases of each one of the selective excitation patterns and another phase of only one of the selective excitation patterns, said another phase being different from the two phases.

18. The system of claim 10, wherein the target is illuminated with three phases of each one of the selective excitation patterns.

19. The system of claim 10, wherein each illumination device generates one of the selective excitation patterns for illuminating the target.

20. An apparatus for performing synthetic aperture optics (SAO) on a target including one or more objects, the apparatus comprising:

a plurality of interference pattern generation devices, each interference pattern generation device configured to generate a pair of light beams that interfere to generate a selective excitation pattern illuminated on the target at a predetermined orientation and a predetermined pitch;

a monolithic half-ring shaped structure holding the plurality of interference pattern generation devices arranged in a half-ring shape;

an optical imaging device configured to optically image the illuminated target at a resolution insufficient to resolve the objects, the illuminated target being imaged by the optical imaging device through a lens included in an imaging path that is physically separated from illumination paths through which the selective excitation pattern from each of the interference pattern generation devices is delivered to the target, the optical image of the illuminated target being further processed using information on the selective excitation pattern to obtain a final image of the illuminated target at a resolution sufficient to resolve the objects.

21. The apparatus of claim 20, wherein the number of the interference pattern generation devices is equal to a number of selective excitation patterns used for performing SAO on the target.

22. The apparatus of claim 20, wherein the interference pattern generation devices are placed substantially symmetrically around the half-ring shape.

23. The apparatus of claim 20, wherein at least one of the interference pattern generation devices includes:

a beam splitter for splitting a first laser beam into the pair of laser beams including a second laser beam and a third laser beam;

a first mirror reflecting the second laser beam onto the target;

a second mirror reflecting the third laser beam; and

a modulator configured to modulate an optical path length of the third laser beam reflected by the second mirror, and

wherein the reflected second laser beam and the reflected, modulated third laser beam interfere to generate the selective excitation pattern on the target.

24. The apparatus of claim 20, wherein at least one of the interference pattern generation devices includes:

a beam splitter for splitting a first laser beam into the pair of laser beams including a second laser beam and a third laser beam;

a first mirror reflecting the second laser beam;

a second mirror further reflecting the second laser beam onto the target;

a third mirror reflecting the third laser beam; and

a modulator configured to modulate an optical path length of the third laser beam reflected by the third mirror, and

wherein the second laser beam and the third laser beam travel substantially equal path lengths before interfering with each other on the target to generate the selective excitation pattern.

25. An apparatus for performing synthetic aperture optics (SAO) on a target including one or more objects, the apparatus comprising:

a plurality of interference pattern generation devices, each interference pattern generation device configured to generate a pair of light beams that interfere to generate a selective excitation pattern illuminating the target at a predetermined orientation and a predetermined pitch, the number of interference pattern generation devices being equal to the number (N) of selective excitation patterns used for performing SAO on the target, and the number (N) of selective excitation patterns corresponding to a number of k-space sampling points in a k-space sampling space in a frequency domain, an extent of the k-space sampling space being substantially proportional to an inverse of the minimum distance (Δ

x) between the objects that is to be resolved by SAO and an inverse of a k-space sampling interval between the k-space sampling points being less than a width (w) of a detected area captured by a pixel of an optical imaging device for optical imaging of the illuminated target; and

a monolithic half-ring shaped structure holding the plurality of interference pattern generation devices arranged in a half-ring shape;

the optical imaging device configured to optically image the illuminated target at a resolution insufficient to resolve the objects, the illuminated target being imaged by the optical imaging device through a lens included in an imaging path that is physically separated from illumination paths through which the selective excitation patterns from the interference pattern generation devices are delivered to the target, the optical image of the illuminated target being further processed using information on the selective excitation patterns to obtain a final image of the illuminated target at a resolution sufficient to resolve the objects.

26. The apparatus of claim 25, wherein the number (N) of selective excitation patterns corresponds to non-uniformly selected ones of the k-space sampling points.

27. The apparatus of claim 25, wherein the interference pattern generation devices are placed substantially symmetrically around the half-ring shape.

28. The apparatus of claim 25, wherein at least one of the interference pattern generation devices includes:

a beam splitter for splitting a first laser beam into the pair of laser beams including a second laser beam and a third laser beam;

a first mirror reflecting the second laser beam onto the target;

a second mirror reflecting the third laser beam; and

a modulator configured to modulate an optical path length of the third laser beam reflected by the second mirror, and

wherein the reflected second laser beam and the reflected, modulated third laser beam interfere to generate the selective excitation pattern on the target.

29. The apparatus of claim 25, wherein at least one of the interference pattern generation devices includes:

a beam splitter for splitting a first laser beam into the pair of laser beams including a second laser beam and a third laser beam;

a first mirror reflecting the second laser beam;

a second mirror further reflecting the second laser beam onto the target;

a third mirror reflecting the third laser beam; and

a modulator configured to modulate an optical path length of the third laser beam reflected by the third mirror, and

wherein the second laser beam and the third laser beam travel substantially equal path lengths before interfering with each other on the target to generate the selective excitation pattern.