Spectral imaging
First Claim
Patent Images
1. A method comprising:
- illuminating a sample to cause the sample to emit radiation, wherein the sample comprises deep tissue supporting a target compound and one or more additional components, and wherein the emitted radiation comprises emission from the target compound and emission from the one or more additional components;
spectrally filtering the emitted radiation with each of a plurality of different spectral weighting functions;
storing an image of the spectrally filtered radiation for each of the spectral weighting functions; and
processing the stored images to construct a deep tissue image of the sample comprising a signal contribution from the target compound and a signal contribution from each of the one or more additional components so that in the deep tissue image, the signal contributions from each of the one or more additional components are reduced relative to the signal contribution from the target compound,wherein processing the stored images comprises constructing the deep tissue image based on the stored images and an emission spectrum for the target compound.
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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.
117 Citations
28 Claims
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1. A method comprising:
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illuminating a sample to cause the sample to emit radiation, wherein the sample comprises deep tissue supporting a target compound and one or more additional components, and wherein the emitted radiation comprises emission from the target compound and emission from the one or more additional components; spectrally filtering the emitted radiation with each of a plurality of different spectral weighting functions; storing an image of the spectrally filtered radiation for each of the spectral weighting functions; and processing the stored images to construct a deep tissue image of the sample comprising a signal contribution from the target compound and a signal contribution from each of the one or more additional components so that in the deep tissue image, the signal contributions from each of the one or more additional components are reduced relative to the signal contribution from the target compound, wherein processing the stored images comprises constructing the deep tissue image based on the stored images and an emission spectrum for the target compound. - 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)
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25. A method comprising:
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providing a plurality of images of spectrally filtered radiation emitted from a sample in response to an illumination, wherein the sample comprises deep tissue supporting a target compound and one or more additional components, wherein the emitted radiation comprises emission from the target compound and emission from the one or more additional components, and wherein each image corresponds to a different spectral weighting function applied to the emitted radiation; and processing the images of the spectrally filtered radiation to construct a deep tissue image of the sample in which a signal contribution from each of the one or more additional components is reduced relative to a signal contribution from the target compound, wherein the processing the images of the spectrally filtered radiation comprises constructing the deep tissue image based on the stored images and an emission spectrum for the target compound.
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26. An apparatus comprising:
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a sample holder configured to hold a sample comprising deep tissue, wherein the deep tissue supports a target compound and one or more additional components; an illumination source configured to illuminate the sample to cause the sample to emit radiation, wherein the emitted radiation comprises emission from the target compound and emission from the one or more additional components; an imaging system configured to image the emitted radiation to a detector; a tunable spectral filter configured to spectrally filter the emitted radiation with each of a plurality of different spectral weighting functions; a detector configured to store an image of the spectrally filtered radiation for each of the spectral weighting functions; and an electronic processor configured to process the stored images to construct a deep tissue image of the sample based on the stored images and an emission spectrum for the target compound, in which a signal contribution from each of the additional components is reduced relative to a signal contribution from the target compound. - View Dependent Claims (27, 28)
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Specification