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US 8,552,402 B2Granted
Super-resolution microscopy system using speckle illumination and array signal processing
AssigneeKorea Advanced Institute of Science and Technology
InventorsJong Chul Ye, Jae Duck Jang
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Abstract
A nano-scale resolution fluorescence microscopy system and a method of obtaining an image using the nano-scale resolution microscopy system, and more particularly, a method and a microscopy system, capable of observing fluorescence probes in high resolution by radiating an irregular diffused light to have an incoherent speckle pattern that has low correlation in an adjacent space are disclosed. According to embodiments of the present invention, a diffraction limit of a fluorescence microscope may be overcome, and a super high resolution image on a nanometer scale may be obtained.
Claims20
A nano-scale resolution fluorescence microscopy system, comprising:
- a speckle pattern generator configured to generate speckle patterns by passing, through a diffuser, a light emitted from at least one coherent light source in at least one color to detect fluorescent emission in at least one color from fluorophores attached to a specimen;
- an illumination optical system to radiate the generated speckle patterns to be exposed on a specimen, by disposing at least one optical lens;
- and
- a fluorescent image optical system configured to detect fluorescence emission images of the specimen excited by the speckle patterns in at least one color, by passing the radiated speckle patterns sequentially through at least one excitation filter and detecting the emission images through at least one emission filter;
- and
- an image reconstruction algorithm configured to sequentially obtain fluorescent emission images by the fluorescent image optical system, and to reconstruct the image at high resolution by detecting temporal profile of fluorescence emission data at every pixel, based on the sequentially obtained images,
- wherein the image reconstruction algorithm is configured to process fluorescence emission images obtained sequentially based on any type of array signal processing algorithms that uses data covariance matrix by calculating the spatial correlation matrix between pixels, and reconstruct images with respect to the fluorophores using the signal and noise subspace structure of the data covariance matrix.
4 more elements
A nano-scale resolution fluorescence microscopy, comprising:
- a speckle pattern generator configured to generate speckle patterns by passing, through a diffuser, a light emitted from at least one coherent light source in at least one color to detect fluorescent emission in at least one color from fluorophores attached to a specimen;
- an illumination optical system configured to radiate the generated speckle patterns to be exposed on a specimen, by disposing at least one optical lens;
- a fluorescent image optical system configured to detect fluorescence emission images of the specimen excited by the speckle patterns in at least one color, by passing the radiated speckle patterns sequentially through at least one excitation filter and detecting the emission images through at least one emission filter;
- and
- an image reconstruction algorithm configured to sequentially obtain fluorescent emission images by the fluorescent image optical system, and to reconstruct the image at high resolution by detecting temporal profile of fluorescence emission data at every pixel, based on the sequentially obtained images,
- wherein the image reconstruction algorithm reconstruct positions of the fluorephores of the target object by exploiting that the measurement data is highly correlated with the numerically calculated forward model.
3 more elements
A method of obtaining an image using a nano-scale resolution microscopy system, the method comprising:
- illuminating coherent light from at least one coherent source to image fluorophores in specimen;
- generating speckle patterns by passing the light emitted from the coherent through a diffuser;
- radiating the generated speckle patterns to be exposed on a specimen, using at least one optical lens;
- photographing a fluorescent light of the speckle patterns, by passing the radiated speckle patterns sequentially through at least one excitation filter and at least one emission filter;
- continuously obtaining images photographed by the fluorescent image optical system;
- and
- restoring the images in high resolution by sensing time dependent data that changes in every pixel, based on the continuously obtained images,
- wherein the restoring of the image in high resolution comprises:
- decomposing a data covariance matrix into the signal and the noise subspaces, and
- reconstructing positions of the fluorephores of the target object by exploiting the orthogonality of a model signal calculated from a fluorophore location with respect to the noise subspace.
7 more elements
A method of obtaining an image using a nano-scale resolution microscopy system, the method comprising:
- illuminating coherent light from at least one coherent source to image fluorophores in specimen;
- generating speckle patterns by passing the light emitted from the coherent through a diffuser;
- radiating the generated speckle patterns to be exposed on a specimen, using at least one optical lens;
- photographing a fluorescent light of the speckle patterns, by passing the radiated speckle patterns sequentially through at least one excitation filter and at least one emission filter;
- continuously obtaining images photographed by the fluorescent image optical system;
- and
- restoring the images in high resolution by sensing time dependent data that changes in every pixel, based on the continuously obtained images,
- wherein the restoring of the image in high resolution comprises:
- decomposing a data covariance matrix into the signal and the noise subspaces, and
- reconstructing positions of the fluorephores of the target object by exploiting that a model signal calculated from a fluorophore location is highly correlated with the signal subspace.
7 more elements
A nano-scale resolution fluorescence microscopy system, comprising:
- a speckle pattern generator configured to generate speckle patterns by passing, through a diffuser, a light emitted from at least one coherent light source in at least one color to detect fluorescent emission in at least one color from fluorophores attached to a specimen;
- an illumination optical system configured to radiate the generated speckle patterns to be exposed on a specimen, by disposing at least one optical lens;
- a fluorescent image optical system configured to detect fluorescence emission images of the specimen excited by the speckle patterns in at least one color, by passing the radiated speckle patterns sequentially through at least one excitation filter and detecting the emission images through at least one emission filter;
- and
- an image reconstruction algorithm configured to sequentially obtain fluorescent emission images by the fluorescent image optical system, and to reconstruct the image at high resolution by detecting temporal profile of fluorescence emission data at every pixel, based on the sequentially obtained images,
- wherein the image reconstruction algorithm is configured to decompose the data covariance matrix into the signal and the noise subspaces, and reconstruct positions of the fluorephores of the target object by exploiting that the signal subspace is highly correlated with the numerically calculated forward model.
3 more elements
Dependencies are inferred from the claim text.