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US 10,335,038 B2Granted
Spectral unmixing for in-vivo imaging
AssigneeXenogen Corporation
InventorsHeng Xu, Bradley W. Rice
Life of this patent
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Abstract
Described herein are systems and methods for spectral unmixing of in vivo light data. The spectral unmixing separates image data according to spectra from multiple internal light sources in an effort to isolate one or more spectrum of interest. The spectral unmixing obtains images with a combination of different and known excitation and emission limits. The spectral unmixing then uses an iterative solution process to separate spectra for the multiple fluorescent light sources, and provides a spectrum and/or a spatial distribution map for at least one of the internal light sources.
Claims31
A method for spectrally unmixing light data corresponding to light emitted from multiple light sources internal to an animal, the method comprising:
- selecting an excitation wavelength band for excitation light provided to the animal;
- selecting an emission wavelength band that limits light wavelengths collected from the animal;
- providing excitation light, into the animal, that is limited to the excitation wavelength band;
- capturing a first image of at least a portion of the animal, where the first image includes light data that is limited in wavelength to the emission wavelength band and corresponds to light emitted by multiple fluorescent light sources internal to the animal;
- changing the excitation wavelength band and/or the emission wavelength band;
- capturing at least one additional image of at least the portion of the animal that uses a different combination of excitation wavelength band and emission wavelength band for each additional image, where each additional image includes light data that corresponds to light emitted by the multiple fluorescent light sources internal to the animal and is limited in wavelength to the emission wavelength band of the different combination;
- and
- unmixing spectra for the multiple fluorescent light sources derived from the first image limited in wavelength to the emission wavelength band and the at least one additional image limited in wavelength to the emission wavelength band to determine a pure spectrum for each light source in the animal using an iterative solution process by a processor,
- wherein said iterative solution process by the processor includes applying both a unimodality constraint that limits the pure spectrum for each light source in the process to having a single peak, and a bandpass constraint that limits the pure spectrum for each light source in the process to within a desired wavelength range.
6 more elements
Logic that is encoded in one or more tangible and non-transitory media for execution by a processor and, when executed, is operable to spectrally unmix spectra corresponding to light emission from multiple light sources internal to an animal, the logic including:
- instructions for selecting an excitation wavelength band for excitation light provided to the animal;
- instructions for selecting an emission wavelength band that limits light wavelengths collected from the animal;
- instructions for providing excitation light, into the animal, that is limited to the excitation wavelength band;
- instructions for capturing a first image of at least a portion of the animal, where the first image includes light data that is limited in wavelength to the emission wavelength band and corresponds to light emitted by multiple fluorescent light sources internal to the animal;
- instructions for changing the excitation wavelength band and/or the emission wavelength band;
- instructions for capturing one or more additional images of at least the portion of the animal, using a different combination of excitation wavelength band and emission wavelength band for each additional image, where each additional image includes light data that corresponds to light emitted by the multiple fluorescent light sources internal to the animal and is limited in wavelength to the emission wavelength band of the different combination;
- and
- instructions for unmixing spectra for the multiple fluorescent light sources derived from the first image limited in wavelength to the emission wavelength band and the at least one additional image limited in wavelength to the emission wavelength band to determine a pure spectrum for each light source in the animal using an iterative solution process,
- wherein said iterative solution process includes applying both a unimodality constraint that limits the pure spectrum for each light source in the process to having a single peak, and a bandpass constraint that limits the pure spectrum for each light source in the process to within a desired wavelength range.
6 more elements
A method for obtaining a three dimensional representation of a light source distribution located inside an animal, the method comprising:
- selecting an excitation wavelength band for excitation light provided to the animal;
- selecting an emission wavelength band that limits light wavelengths collected from the animal;
- providing excitation light, into the animal, that is limited to the excitation wavelength band;
- capturing a first image of at least a portion of the animal, where the first image includes light data that is limited in wavelength to the emission wavelength band and corresponds to light emitted by multiple fluorescent light sources internal to the animal;
- changing the excitation wavelength band and/or the emission wavelength band;
- capturing one or more additional images of at least the portion of the animal, using a different combination of excitation wavelength band and emission wavelength band for each additional image, wherein each additional image includes light data that corresponds to light emitted by the multiple fluorescent light sources internal to the animal and is limited in wavelength to the emission wavelength band of the different combination;
- using an iterative solution process by a processor, unmixing spectra for the multiple fluorescent light sources derived from the first image limited in wavelength to the emission band and the at least one additional image limited in wavelength to the emission band to provide a pure spectrum and spatial distribution map for one of the multiple fluorescent light sources;
- obtaining a three dimensional representation of a surface portion of the animal;
- combining the spatial distribution map and the three dimensional representation of the surface portion of the animal to generate a three-dimensional distribution of light emission from the surface portion of the animal corresponding to the one fluorescent light source;
- and
- determining a three-dimensional representation of a distribution of the one fluorescent light source internal to the animal using the three-dimensional distribution of light emission from the surface portion of the animal corresponding to the one fluorescent light source.
8 more elements
A method for spectrally unmixing light data corresponding to light emitted from multiple light sources internal to an animal, the method comprising:
- selecting an emission wavelength band that limits light wavelengths collected from the animal;
- capturing a first image of at least a portion of the animal, where the first image includes light data that is limited in wavelength to the emission wavelength band and corresponds to light emitted by the multiple light sources internal to the animal;
- changing the emission wavelength band;
- capturing one or more additional images of at least the portion of the animal, using a different emission wavelength band for each additional image, where each additional image includes light data that corresponds to light emitted by the multiple light sources internal to the animal and is limited in wavelength to the different emission wavelength band;
- and
- using an iterative solution process by a processor, unmixing spectra for the multiple light sources derived from the first image limited in wavelength to the emission wavelength band and the at least one additional image limited in wavelength to the emission wavelength band to determine a pure spectrum for each light source in the animal,
- wherein the iterative solution process implements a multivariate curve resolution and iteratively solves for the pure spectrum of each of the multiple light sources using finishing criteria;
- and
- wherein said iterative solution process by the processor includes applying both a unimodality constraint that limits the pure spectrum for each light source in the process to having a single peak, and a bandpass constraint that limits the pure spectrum for each light source in the process to within a desired wavelength range.
6 more elements
An imaging system for imaging an animal, the imaging system comprising:
- an imaging chamber that includes
- a set of walls and a door that enclose an interior cavity,
- a stage configured to support the animal within the interior cavity,
- a camera, and
- a fluorescent excitation source;
- and
- a processing system including a processor and memory, the memory including:
- instructions for selecting an excitation wavelength band for excitation light provided to the animal,
- instructions for selecting an emission wavelength band that limits light spectra collected from the animal,
- instructions for obtaining one or more fluorescent images using the camera, and
- instructions for unmixing spectra for the multiple fluorescent light sources derived from the one or more fluorescent images to determine a pure spectrum for each fluorescent light source using an iterative solution process,
- wherein said iterative solution process comprises applying a unimodality constraint that limits the pure spectrum for each light source to having a single peak, and a bandpass constraint that limits the pure spectrum for each light source to within a desired wavelength range.
9 more elements
Dependencies are inferred from the claim text.