Spectral imaging of biological samples
AssigneeCambridge Research & Instrumentation, Inc.
InventorsRichard Levenson, Paul J. Cronin, Kirk William Gossage, Clifford C. Hoyt
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Abstract
The invention features a method including: (i) providing spectrally resolved information about light coming from different spatial locations in a sample comprising deep tissue in response to an illumination of the sample, wherein the light includes contributions from different components in the sample; (ii) decomposing the spectrally resolved information for each of at least some of the different spatial locations into contributions from spectral estimates associated with at least some of the components in the sample; and (iii) constructing a deep tissue image of the sample based on the decomposition to preferentially show a selected one of the components.
Claims24
A method for spectrally unmixing light data corresponding to light emitted from multiple different fluorescent 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 the multiple different fluorescent light sources in the emission wavelength band 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 different fluorescent light sources internal to the animal and is limited in wavelength to the emission wavelength band of the different combination;
- and
- using an iterative solution process, unmixing spectra for the multiple fluorescent light sources internal to the animal the light data in the images to provide a spectrum for each light source of the multiple different fluorescent light sources,
- wherein each spectrum provides wavelength-dependent excitation and/or emission information about the corresponding fluorescent light source.
Logic encoded in one or more non-transitory storage devices for execution and, when executed, operable to spectrally unmix light data corresponding to light emitted from multiple different fluorescent 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 the multiple different fluorescent light sources in the emission wavelength band 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 different 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 the light data in the images spectra, using an iterative solution process, for the multiple fluorescent light sources internal to the animal to provide a spectrum for each of the multiple different light fluorescent sources,
- light source wherein each spectrum provides wavelength-dependent excitation and/or emission information about the corresponding fluorescent light source.
A method for spectrally unmixing light data corresponding to light emitted from multiple different 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 different light sources in the emission wavelength band 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 different light sources internal to the animal and is limited in wavelength to the different emission wavelength band;
- and
- using an iterative solution process, unmixing the light data in the images spectra for the multiple light sources internal to the animal to provide an emission spectrum for each of the multiple different light sources light source, wherein the iterative solution process implements a multivariate curve resolution and iteratively solves for each emission spectrum a spectrum of each of the multiple light sources using finishing criteria,
- wherein each emission spectrum provides wavelength-dependent emission information about the corresponding light source.
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 non-transitory 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 wavelengths collected from the animal,
- instructions for obtaining one or more fluorescent images using the camera, and
- instructions for unmixing the fluorescent images spectra, using an iterative solution process, for the multiple fluorescent light sources internal to the animal to provide a spectrum for each of multiple different fluorescent light source sources internal to the animal,
- wherein each spectrum provides wavelength-dependent excitation and/or emission information about the corresponding fluorescent light source.
Dependencies are inferred from the claim text.