Multi-well fourier ptychographic and fluorescence imaging
First Claim
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1. An imaging device for bright field Fourier ptychographic imaging and fluorescence imaging, the imaging device comprising:
- a first fluorescence illumination source configured to provide excitation light of a first range of wavelengths to a transparent well;
an optical system having a pair of objectives, the pair of objectives comprising a first objective and a second objective positioned oppositely; and
an image sensor configured to receive light propagated through the optical system from the transparent well, the image sensor further configured to acquire a sequence of uniquely illuminated intensity measurements of light passing through a sample being imaged during operation in the well based on sequential illumination at different illumination angles from a variable illumination source, the image sensor further configured to acquire a first fluorescence image of the sample based on light emitted by the sample in response to excitation light of the first range of wavelengths, wherein the first objective is configured to collect light from the transparent well and the second objective is configured to focus light to the image sensor.
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Abstract
Certain aspects pertain to multi-well systems, devices, and methods of Fourier ptychographic and fluorescence imaging.
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Citations
36 Claims
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1. An imaging device for bright field Fourier ptychographic imaging and fluorescence imaging, the imaging device comprising:
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a first fluorescence illumination source configured to provide excitation light of a first range of wavelengths to a transparent well; an optical system having a pair of objectives, the pair of objectives comprising a first objective and a second objective positioned oppositely; and an image sensor configured to receive light propagated through the optical system from the transparent well, the image sensor further configured to acquire a sequence of uniquely illuminated intensity measurements of light passing through a sample being imaged during operation in the well based on sequential illumination at different illumination angles from a variable illumination source, the image sensor further configured to acquire a first fluorescence image of the sample based on light emitted by the sample in response to excitation light of the first range of wavelengths, wherein the first objective is configured to collect light from the transparent well and the second objective is configured to focus light to the image sensor. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 34)
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16. A system for Fourier ptychographic imaging and fluorescence imaging, the system comprising:
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a body configured to receive a multi-well plate; imaging devices arranged for one to one correspondence with transparent wells in the multi-well plate, each imaging device comprising; a first fluorescence illumination source configured to provide excitation light of a first range of wavelengths to the corresponding well; an optical system having a pair of objectives, the pair of objectives comprising a first objective and a second objective positioned oppositely; and an image sensor for capturing intensity measurements based on light received from the corresponding well, wherein the first objective is configured to collect light from the corresponding well and the second objective is configured to focus light to the image sensor; and a processor configured to generate an improved resolution brightfield image of each sample being imaged in a respective well during operation, the processor configured to generate the improved brightfield image using Fourier ptychographic reconstruction based on a sequence of uniquely illumination intensity measurements acquired during sequential illumination at different illumination angles by a variable illumination source; and wherein the system is also configured to generate a first fluorescence image of each sample being imaged based on light emitted by the sample in response to receiving excitation light of the first range of wavelengths. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. An imaging method, comprising:
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calibrating positions of light sources of a variable illumination source to locations of image sensors receiving light from wells in a multi-well plate; illuminating the multi-well plate with excitation light of a first range of wavelengths; for each well in the multi-well plate, acquiring a first fluorescence image of a sample being imaged during operation in the well; sequentially illuminating the multi-well plate with plane wave illumination at a plurality of illumination angles using a variable illumination source; for each well in the multi-well plate, acquiring, using an image sensor, a sequence of uniquely illuminated intensity measurements from light passing through the sample being imaged in the corresponding well based on the sequential illumination at the plurality of illumination angles; and for each well in the multi-well plate, reconstructing with a Fourier ptychography reconstruction process an improved resolution brightfield image of the sample being imaged based on the sequence of uniquely illuminated intensity measurements. - View Dependent Claims (32)
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33. An imaging method, comprising:
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sequentially illuminating a multi-well plate with plane wave illumination at a plurality of illumination angles using a variable illumination source; for each well in the multi-well plate, acquiring, using an image sensor, a sequence of intensity measurements from light passing through the well based on sequential illumination at the plurality of illumination angles; for each well in the multi-well plate, reconstructing, using a Fourier ptychography reconstruction process, an improved resolution brightfield image using the acquired sequence of intensity measurements; illuminating the multi-well plate with excitation light of a first range of wavelengths; for each well in the multi-well plate, acquiring, using the image sensor, a first fluorescence image based on emissions resulting from the illumination by excitation light of the first range of wavelengths; illuminating the multi-well plate with excitation light of a second range of wavelengths; and for each well in the multi-well plate, acquiring, using the image sensor, a second fluorescence image based on emissions resulting from the illumination by excitation light of the second range of wavelengths and overlaying the first and second fluorescence images to generate a multicolor fluorescence image.
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35. An imaging method, comprising:
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sequentially illuminating a multi-well plate with plane wave illumination at a plurality of illumination angles using a variable illumination source; for each well in the multi-well plate, collecting light using a first objective and focusing the collected light to an imaging sensor using a second objective, the first objective and the second objective positioned oppositely; for each well in the multi-well plate, acquiring, using the image sensor, a sequence of intensity measurements from light passing through the well based on the sequential illumination at the plurality of illumination angles; reconstructing an improved resolution brightfield image using a Fourier ptychography reconstruction process and based on the acquired sequence of intensity measurements; illuminating the multi-well plate with excitation light; and for each well in the multi-well plate, acquiring, using the image sensor, a fluorescence image. - View Dependent Claims (36)
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Specification