Spectral imaging
First Claim
1. A method for analyzing spectrally resolved information about light coming from different spatial locations of a biological sample in response to an illumination of the sample, wherein the light includes contributions from different components in the sample, said method comprising using one or more electronic processors to perform the following:
- deriving an estimate of a pure spectrum for at least a first one of the components in the sample from at least part of the spectrally resolved information corresponding to a first set of one or more of the different spatial locations and a spectral estimate of a pure spectrum for a second one of the components, wherein each pure spectrum estimate corresponds to the spectrally resolved information that would result if only the component corresponding to that estimate contributes to the light and wherein each pure spectrum estimate comprises a plurality of values corresponding to different spectral weighting functions;
decomposing at least some of the spectrally resolved information for each of at least some of the different spatial locations into a contribution from the spectral estimate of the pure spectrum for at least a selected one of the components in the sample; and
constructing an image of the sample based on the decomposition to preferentially show the selected one of the components.
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Accused Products
Abstract
Apparatus and methods are provided for the imaging of structures in deep tissue within biological specimens, using spectral imaging to provide highly sensitive detection. By acquiring data that provides a plurality of images of the sample with different spectral weightings, and subsequent spectral analysis, light emission from a target compound is separated from autofluorescence in the sample. With the autofluorescence reduced or eliminated, an improved measurement of the target compound is obtained.
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Citations
31 Claims
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1. A method for analyzing spectrally resolved information about light coming from different spatial locations of a biological sample in response to an illumination of the sample, wherein the light includes contributions from different components in the sample, said method comprising using one or more electronic processors to perform the following:
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deriving an estimate of a pure spectrum for at least a first one of the components in the sample from at least part of the spectrally resolved information corresponding to a first set of one or more of the different spatial locations and a spectral estimate of a pure spectrum for a second one of the components, wherein each pure spectrum estimate corresponds to the spectrally resolved information that would result if only the component corresponding to that estimate contributes to the light and wherein each pure spectrum estimate comprises a plurality of values corresponding to different spectral weighting functions; decomposing at least some of the spectrally resolved information for each of at least some of the different spatial locations into a contribution from the spectral estimate of the pure spectrum for at least a selected one of the components in the sample; and constructing an image of the sample based on the decomposition to preferentially show the selected one of the components. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. A method for analyzing a set of images of spectrally filtered radiation emitted from a biological sample in response to an illumination, the method comprising:
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providing the set of images, wherein the sample comprises a component supporting a target compound and the emitted radiation comprises emission from the target compound and emission from one or more additional components in the sample, and wherein each image corresponds to a different spectral weighting function for a common set of pixels; and using one or more electronic processors to process the images of the spectrally filtered radiation to construct an output image of the sample in which signal from the additional components is reduced relative to signal from the target compound, wherein the processing comprises calculating a remainder spectrum for one or more pixels in the set of images based on an estimate for a pure spectrum of at least one of the components; wherein, for each of the at least one of the components, the estimate for the pure spectrum corresponds to the spectrally filtered radiation that would result if only that component contributed to the emitted radiation, and the estimate for the pure spectrum comprises a plurality of values corresponding to different spectral weighting functions; and wherein for each of the one or more pixels, the remainder spectrum is calculated according to
Ra(λ
)=S(λ
)−
aF(λ
)where Ra(λ
) is the remainder spectrum, S(λ
) is the spectrally filtered radiation corresponding to the pixel, F(λ
) is the estimate of the pure spectrum of the at least one of the components, a is a parameter, and λ
is a wavelength of the spectrally filtered radiation.
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Specification