Correcting for aberrations in incoherent imaging systems using fourier ptychographic techniques

Patent metadata

Field	Value
Publication number	US-10684458-B2
Application number	US-201615068389-A
Country	US
Kind code	B2
Filing date	Mar 11, 2016
Priority date	Mar 13, 2015
Publication date	Jun 16, 2020
Grant date	Jun 16, 2020

Abstract

Aberration-corrected incoherent imaging methods and systems that can acquire a sequence of coherent images and an incoherent image of a specimen, implement an embedded pupil function recovery process in junction with Fourier ptychographic technique to construct an improved resolution image and pupil function of the imaging system using the sequence of coherent images, determine an optical transfer function based on the estimated pupil function, and remove the aberration from the incoherent image using a deconvolution process to generate an aberration-corrected incoherent image.

First claim

What is claimed is:

1. An aberration-corrected fluorescence imaging method implemented by an imaging system having an optical system and an image sensor configured to receive radiation transmitted by the optical system, the method comprising:
acquiring, using the image sensor, a sequence of coherent images of a specimen while spatially coherent plane wave illumination from different angles is sequentially provided incident the specimen, wherein each image of the sequence of coherent images is acquired while coherent plane wave illumination is provided incident the specimen from one of the angles;
illuminating the specimen with excitation light configured to cause fluorescence emissions;
receiving, at collection optics of the optical system, light including fluorescence emissions issuing from the specimen;
passing fluorescence emissions and substantially blocking excitation light using an emission filter of the optical system;
acquiring, using the image sensor, a fluorescence image of the specimen based on fluorescence emissions passed from the optical system;
implementing an embedded pupil function recovery process to use the acquired sequence of coherent images to estimate a pupil function of the imaging system and construct an improved resolution image, wherein construction of the improved resolution image and estimation of the pupil function occur simultaneously;
determining an optical transfer function of the imaging system based on the estimated pupil function;
removing aberration from the acquired fluorescence image using a non-blind deconvolution process with one or more regularization parameters to generate an aberration-corrected fluorescence image, wherein the non-blind deconvolution process uses the optical transfer function of the imaging system determined from the pupil function estimated using the acquired sequence of coherent images; and
causing the aberration-corrected fluorescence image to be stored on a non-transitory storage medium,
wherein the improved resolution image and the aberration-corrected fluorescence image are co-localized, and wherein the aberration-corrected fluorescence image is a monochromatic fluorescence image.

2. The aberration-corrected fluorescence imaging method of claim 1 , further comprising converting the monochromatic fluorescence image into a color fluorescence image.

3. The aberration-corrected fluorescence imaging method of claim 1 , further comprising:
dividing a field-of-view of each of the acquired sequence of coherent images into a plurality of tile regions;
generating a sequence of tile images for each tile region;
generating a fluorescence tile image for each tile region; and
for each tile region, implementing the embedded pupil function recovery process to construct an improved resolution tile image and estimate a tile pupil function using the generated sequence of tile images.

4. The aberration-corrected fluorescence imaging method of claim 3 , wherein the improved resolution tile image is constructed for each tile in parallel and the tile pupil function is estimated for each tile region in parallel.

5. The aberration-corrected fluorescence imaging method of claim 3 , further comprising:
for each tile region, determining a tile optical transfer function based on the estimated tile pupil function;
for each tile region, removing aberration from the fluorescence tile image to generate an aberration-corrected fluorescence tile image using the non-blind deconvolution process and with the determined tile optical transfer function; and
generating the aberration-corrected fluorescence image by combining the aberration-corrected fluorescence tile images for the plurality of tile regions.

6. The aberration-corrected fluorescence imaging method of claim 1 , wherein implementing the embedded pupil function recovery process to use the acquired sequence of coherent images to construct the improved resolution image comprises recovering amplitude and phase data of the improved resolution image.

7. The aberration-corrected fluorescence imaging method of claim 1 , wherein the illumination angles are oblique angles to a surface of the specimen being imaged.

8. The aberration-corrected fluorescence imaging method of claim 1 , further comprising:
acquiring at least one additional fluorescence image of the specimen; and
removing the aberration from the at least additional fluorescence image to generate aberration-corrected fluorescence images.

9. The aberration-corrected fluorescence imaging method of claim 8 ,
wherein the at least one additional fluorescence image is monochromatic, and
further comprising overlaying the monochromatic aberration-corrected fluorescence images to generate a multi-color fluorescence image of the specimen.

10. The aberration-corrected fluorescence imaging method of claim 1 , wherein the fluorescence image is acquired before the sequence of coherent images is acquired.

11. The aberration-corrected fluorescence imaging method of claim 1 , wherein the fluorescence image is acquired after implementing the embedded pupil function recover process or determining the optical transfer function based on the estimated pupil function.

12. An imaging system comprising:
a variable coherent light source configured to sequentially illuminate a specimen with spatially coherent plane wave illumination from different oblique angles;
an excitation light source configured to provide excitation light configured to cause fluorescence emissions from the specimen;
an optical system with collection optics for collecting light issuing from the specimen, wherein the optical system includes an emission filter for passing fluorescence emissions and substantially blocking excitation light, the optical system configured to propagate light to one or more image sensors;
the one or more image sensors configured to acquire a sequence of coherent images of the specimen while the variable coherent light source sequentially illuminates the specimen with coherent plane wave illumination from different oblique angles, the one or more image sensors further configured to acquire a fluorescence image of the specimen while the excitation light source provides excitation light incident the specimen; and
one or more processors in electrical communication with the one or more image sensors to receive image data of the sequence of coherent images and the fluorescence image, the one or more processors also configured to implement instructions stored in memory to:
estimate a pupil function of the imaging system and construct an improved resolution image using an embedded pupil function recovery process with the image data of the acquired sequence of coherent images, wherein construction of the improved resolution image and estimation of the pupil function occur simultaneously;
determine an optical transfer function of the imaging system based on the estimated pupil function;
generate an aberration-corrected fluorescence image by removing aberration from the acquired fluorescence image using a non-blind deconvolution process with one or more regularization parameters, wherein the non-blind deconvolution process uses the optical transfer function of the imaging system determined from the pupil function estimated using the acquired sequence of coherent images, wherein the improved resolution image and the aberration-corrected fluorescence image are co-localized, and wherein the aberration-corrected fluorescence image is a monochromatic fluorescence image; and
cause the aberration-corrected fluorescence image to be stored on a non-transitory storage medium.

13. The imaging system of claim 12 , wherein the excitatio

Assignees

• California Inst Of Techn

Inventors

• Chung Jaebum
• Yang Changhuei

Classifications

• G02B21/0072 Primary

  details concerning resolution or correction, including general design of CSOM objectives · CPC title

• G02B21/0076 Primary

  arrangements using fluorescence or luminescence · CPC title

• G02B21/0032

  Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers (G02B21/0036 - G02B21/008; means for illumination of specimens in general G02B21/06) · CPC title

• G02B21/0056

  based on optical coherence, e.g. phase-contrast arrangements, interference arrangements · CPC title

• G02B21/008

  Details of detection or image processing, including general computer control · CPC title