Systems and methods for rapid wide field illumination scanning for in vivo small animal fluorescence tomographic imaging

A system for fast scanning of excitation light from a plurality of excitation sources over a wide field of view for tomographic imaging of a plurality of subjects positioned across an object plane, the system comprising:

• (a) a plurality of excitation sources, wherein:
• each excitation source is operable to emit a respective beam of excitation light,
• a first excitation source is aligned to direct its respective beam of excitation light along a first optical path from an output of the first excitation source to a galvanometer optical scanner comprising one or more rotating galvanometer mirrors, and
• each excitation source other than the first excitation source is aligned to direct its respective beam of excitation light along a respective optical path from its output to the galvanometer optical scanner at a corresponding offset angle with respect to the first optical path;
• (b) the galvanometer optical scanner, wherein the galvanometer optical scanner is aligned and operable to, for each excitation source, direct, during an exposure of the plurality of subjects to the respective beam of excitation light, the respective beam of excitation light to a respective plurality of discrete excitation locations within a respective scan region of the object plane via reflection by the one or more rotating galvanometer mirrors, such that as the one or more galvanometer mirrors are rotated, the respective beam of excitation light is scanned across the respective scan region, thereby providing for illumination of the plurality of subjects positioned across the object plane,
• wherein the respective plurality of discrete excitation locations comprise a plurality of multi-subject sets of discrete excitation locations, and wherein each of the multi-subject sets of discrete excitation locations comprises a first discrete excitation location corresponding to a first location of a first subject of the plurality of subjects and a second discrete excitation location corresponding to a second location of a second subject of the plurality of subjects, and
• wherein the plurality of subjects are statically fixed to the object plane as the one or more galvanometer mirrors are rotated;
• (c) one or more detectors aligned and operable to detect, during the exposure of the plurality of subjects to the respective beam of excitation light, fluorescent light emitted from one or more fluorescent species of the plurality of subjects as a result of excitation by the respective beam of excitation light at the first discrete excitation location and at the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations;
• (d) a processor;
• and
• (e) a memory having instructions stored thereon, wherein the instructions, when executed by the processor cause the processor to:
• receive and/or access data corresponding to the detected fluorescent light;
• and
• obtain one or more tomographic images of the plurality of subjects using the data corresponding to the detected fluorescent light, wherein the one or more tomographic images comprises a multi-subject tomographic image that represents the fluorescent light emitted from the plurality of subjects at the first discrete excitation location and the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations.

A method for fast scanning of excitation light over a wide field of view for tomographic imaging of a plurality of subjects positioned across an object plane, the method comprising:

• (a) illuminating the plurality of subjects with a beam of excitation light from an excitation source, wherein the excitation source is aligned to direct the beam of excitation light along an optical path from an output of the excitation source to a galvanometer optical scanner comprising one or more rotating galvanometer mirrors, wherein the galvanometer optical scanner is aligned and operable to direct, during an exposure of the plurality of subjects to the beam of excitation light, the beam of excitation light to a plurality of discrete excitation locations within a scan region of the object plane via reflection by the one or more rotating galvanometer mirrors, such that as the one or more galvanometer mirrors are rotated, the beam of excitation light is scanned across the scan region, thereby providing for illumination of the plurality of subjects positioned across the object plane,
• wherein the plurality of discrete excitation locations comprise a plurality of multi-subject sets of discrete excitation locations, and wherein each of the multi-subject sets of discrete excitation locations comprises a first discrete excitation location corresponding to a first location of a first subject of the plurality of subjects and a second discrete excitation location corresponding to a second location of a second subject of the plurality of subjects, and
• wherein the plurality of subjects are within the scan region and are statically fixed to the object plane as the one or more galvanometer mirrors are rotated;
• (b) detecting, with one or more detectors and during the exposure of the plurality of subjects to the beam of excitation light, fluorescent light emitted from a fluorescent species within the plurality of subjects as a result of excitation by the beam of excitation light at the first discrete excitation location and at the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations;
• (c) receiving and/or accessing, by a processor of a computing device, data corresponding to the detected fluorescent light;
• and
• (d) obtaining, by the processor, one or more tomographic images of the plurality of subjects using the data corresponding to the detected fluorescent light, wherein the one or more tomographic images comprises a multi-subject tomographic image that represents the fluorescent light emitted from the plurality of subjects at the first discrete excitation location and the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations.

A method for fast scanning of excitation light from a plurality of excitation sources over a wide field of view for tomographic imaging of a plurality of subjects positioned across an object plane, the method comprising:

• (a) illuminating the plurality of subjects with at least one beam of excitation light emitted by a respective one of a plurality of excitation sources, wherein:
• (i) a first excitation source is aligned to direct its respective beam of excitation light along a first optical path from the first excitation source to a galvanometer optical scanner comprising one or more rotating galvanometer mirrors,
• (ii) each excitation source other than the first excitation source is aligned to direct its respective beam of excitation light along an respective optical path towards the galvanometer optical scanner at a corresponding offset angle with respect to the first optical path from the first excitation source to the galvanometer optical scanner,
• (iii) the galvanometer optical scanner is aligned and operable to, for each excitation source, direct, during an exposure of the plurality of subjects to the respective beam of excitation light, the respective beam of excitation light to a respective plurality of discrete excitation locations within a respective scan region of the object plane via reflection by the one or more rotating galvanometer mirrors, such that as the one or more galvanometer mirrors are rotated, the respective beam of excitation light is scanned across the respective scan region, thereby providing for illumination of the plurality of subjects positioned across the object plane,
• wherein the respective plurality of discrete excitation locations comprise a plurality of multi-subject sets of discrete excitation locations, and wherein each of the multi-subject sets of discrete excitation locations comprises a first discrete excitation location corresponding to a first location of a first subject of the plurality of subjects and a second discrete excitation location corresponding to a second location of a second subject of the plurality of subjects, and
• wherein the plurality of subjects are within the scan region and are statically fixed to the object plane as the one or more galvanometer mirrors are rotated;
• (b) detecting, with one or more detectors and during the exposure of the plurality of subjects to the respective beam of excitation light, fluorescent light emitted from one or more fluorescent species of the plurality of subjects as a result of excitation by the respective beam of excitation light at the first discrete excitation location and at the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations;
• (c) receiving and/or accessing, by a processor of a computing device, data corresponding to the detected fluorescent light;
• and
• (d) obtaining, by the processor, one or more tomographic images of the plurality of subjects using the data corresponding to the detected fluorescent light, wherein the one or more tomographic images comprises a multi-subject tomographic image that represents the fluorescent light emitted from the plurality of subjects at the first discrete excitation location and the second discrete excitation location of one of the plurality of multi-subject sets of discrete excitation locations.