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

• US 10,429,665 B2
• Filed: 08/24/2017
• Issued: 10/01/2019
• 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:
  US15/685,982
• Publication Number:
  US 20180136481A1
• Time in Patent Office: 768 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, wherein the target is illuminated with two or three phases of each one of the selective excitation patterns;

optically imaging the illuminated target at a first resolution, wherein the optical imaging is from illuminating the target with the two or three phases of each one of the selective excitation patterns; 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 second resolution higher than the first resolution.

2. 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.

3. 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 a minimum distance (Δ

x) 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 predetermined width (w) of a detected area captured by a pixel of an optical imaging system for optical imaging of the illuminated target.

4. The method of claim 3, 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.

5. The method of claim 3, 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 predetermined width (w) of the detected area captured by the pixel of said optical imaging system.

6. 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 a 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 is determined based upon at least one of a plurality of parameters of the optical imaging system, including a predetermined width (w) of a detected area captured by a pixel of an optical imaging system for optical imaging of the illuminated target, a wavelength of light from the illuminated target, and a numerical aperture of a lens through which said light is passed through in the optical imaging system.

8. 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.

9. The method of claim 1, wherein the illuminated target is imaged by the optical imaging system through a lens included in an imaging path that is physically separated from an illumination path through which the N selective excitation patterns are delivered to the target.

10. 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 modules (IPGMs), each IPGM 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, each IPGM further configured to generate two or three phases of the corresponding selective excitation pattern, the number of IPGMs being equal to a predetermined number (N) of selective excitation patterns used for performing SAO on the target, and the predetermined 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; and

an optical imaging module configured to optically image the illuminated target at a first resolution, the optical imaging module further configured to optically image the illuminated target from illuminating the target with the two or three phases of the selective excitation pattern from each of the plurality of IPGMs, 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 second resolution higher than the first resolution.

11. The apparatus of claim 10, wherein the optical imaging module is configured to optically image the illuminated target based on measurements obtained from illuminating the target with two phases of each one of the selective excitation patterns from the plurality of IPGMs and another phase of only one of the selective excitation patterns from one IPGM, said another phase being different from the two phases.

12. The apparatus of claim 10, wherein at least one of the IPGM 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, and wherein the two or three phases of the selective excitation pattern of the at least one IPGM is generated by adjusting the optical path length of the third laser beam.

13. The apparatus of claim 10, wherein the illuminated target is imaged by the optical imaging module through a lens included in an imaging path that is physically separated from an illumination path through which the selective excitation pattern from each of the IPGMs is delivered to the target.

14. The apparatus of claim 10, further comprising a monolithic half ring structure holding the plurality of IPGMs arranged in a half ring shape.

15. 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 modules (IPGMs), each IPGM 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, each IPGM further configured to generate two or three phases of the selective excitation pattern;

a monolithic half-ring shaped structure holding the plurality of IPGMs arranged in a half-ring shape;

an optical imaging module configured to optically image the illuminated target at a first resolution, the optical imaging module further configured to optically image the illuminated target from illuminating the target with the two or three phases of the selective excitation pattern from each of the plurality of IPGMs, 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 second resolution higher than the first resolution.

16. The apparatus of claim 15, wherein the optical imaging module is configured to optically image the illuminated target based on measurements obtained from illuminating the target with two phases of each one of the selective excitation patterns from the plurality of IPGMs and another phase of only one of the selective excitation patterns from one IPGM, said another phase being different from the two phases.

17. The apparatus of claim 15, wherein at least one of the IPGM 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, and wherein the two or three phases of the selective excitation pattern of the at least one IPGM is generated by adjusting the optical path length of the third laser beam.

18. The apparatus of claim 15, wherein the illuminated target is imaged by the optical imaging module through a lens included in an imaging path that is physically separated from an illumination path through which the selective excitation pattern from each of the IPGMs is delivered to the target.