Spectral imaging of deep tissue
First Claim
Patent Images
1. A method comprising:
- illuminating a sample to cause the sample to emit radiation, wherein the sample is a living mammal comprising deep tissue supporting a target compound, wherein the emitted radiation comprises emission from the target compound and emission from one or more other components in the sample, wherein the emission from the other components of the sample comprises autofluorescence from one or more layers of tissue in the sample different from a layer of tissue comprising the deep tissue, and wherein the deep tissue is subdermal tissue;
spectrally filtering the emitted radiation with each of at least four 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 in which signal from the other compounds is reduced relative to signal from the target compound, wherein processing the stored images comprises constructing the deep tissue image based on a weighted superposition of information in the stored images and at least one emission spectrum for the other components in the sample.
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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.
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Citations
34 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 is a living mammal comprising deep tissue supporting a target compound, wherein the emitted radiation comprises emission from the target compound and emission from one or more other components in the sample, wherein the emission from the other components of the sample comprises autofluorescence from one or more layers of tissue in the sample different from a layer of tissue comprising the deep tissue, and wherein the deep tissue is subdermal tissue; spectrally filtering the emitted radiation with each of at least four 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 in which signal from the other compounds is reduced relative to signal from the target compound, wherein processing the stored images comprises constructing the deep tissue image based on a weighted superposition of information in the stored images and at least one emission spectrum for the other components in the sample. - 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)
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26. A method comprising:
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providing at least four images of spectrally filtered radiation emitted from a sample that is a living mammal in response to an illumination, wherein the sample comprises deep tissue supporting a target compound, wherein the emitted radiation comprises emission from the target compound and emission from one or more other components in the sample, wherein the emission from the other components of the sample comprises autofluorescence from one or more layers of tissue in the sample different from a layer of tissue comprising the deep tissue, wherein the deep tissue is subdermal tissue, and wherein each image corresponds to a different spectral weighting function; and processing the images of the spectrally filtered radiation to construct a deep tissue image of the sample in which signal from the other compounds is reduced relative to signal from the target compound, and wherein processing the images comprises constructing the deep tissue image based on a weighted superposition of information in the stored images and at least one emission spectrum for the other components in the sample. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34)
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Specification