3D imaging of live cells with ultraviolet radiation
First Claim
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1. A method for 3D imaging of a biological object in an optical tomography system comprising:
- moving a biological object relative to a microscope objective to present varying angles of view;
illuminating the entire biological object;
axially scanning the entire biological object with the microscope objective at a fixed viewpoint through the thickness of the biological object where an axial scan is performed at each of said varying angles of view;
sensing a first radiation bandwidth transmitted through the entire biological object and the microscope objective with an ultraviolet camera during axially scanning, wherein the first radiation bandwidth has a first spectral range between 150 nm and 390 nm to produce a first plurality of pseudo-projection images of the specimen onto the ultraviolet camera;
sensing a second radiation bandwidth within a second spectral range which is also transmitted through the entire biological object and the microscope objective to the ultraviolet camera during axially scanning, wherein the second spectral range is also between 150 nm and 390 nm, but different than the first spectral range, to produce a second plurality of pseudo-projection images of the specimen onto the ultraviolet camera;
wherein each axial scan is performed during a single exposure of the ultraviolet camera, and wherein each pseudo-projection image thus formed includes an integration of a range of focal plane images; and
reconstructing the plurality of pseudoprojection images to form a reconstructed 3D image.
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Abstract
A method for 3D imaging of cells in an optical tomography system includes moving a biological object relatively to a microscope objective to present varying angles of view. The biological object is illuminated with radiation having a spectral bandwidth limited to wavelengths between 150 nm and 390 nm. Radiation transmitted through the biological object and the microscope objective is sensed with a camera from a plurality of differing view angles. A plurality of pseudoprojections of the biological object from the sensed radiation is formed and the plurality of pseudoprojections is reconstructed to form a 3D image of the cell.
124 Citations
63 Claims
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1. A method for 3D imaging of a biological object in an optical tomography system comprising:
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moving a biological object relative to a microscope objective to present varying angles of view; illuminating the entire biological object; axially scanning the entire biological object with the microscope objective at a fixed viewpoint through the thickness of the biological object where an axial scan is performed at each of said varying angles of view; sensing a first radiation bandwidth transmitted through the entire biological object and the microscope objective with an ultraviolet camera during axially scanning, wherein the first radiation bandwidth has a first spectral range between 150 nm and 390 nm to produce a first plurality of pseudo-projection images of the specimen onto the ultraviolet camera; sensing a second radiation bandwidth within a second spectral range which is also transmitted through the entire biological object and the microscope objective to the ultraviolet camera during axially scanning, wherein the second spectral range is also between 150 nm and 390 nm, but different than the first spectral range, to produce a second plurality of pseudo-projection images of the specimen onto the ultraviolet camera; wherein each axial scan is performed during a single exposure of the ultraviolet camera, and wherein each pseudo-projection image thus formed includes an integration of a range of focal plane images; and reconstructing the plurality of pseudoprojection images to form a reconstructed 3D image. - 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, 26, 27, 28, 29)
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30. An optical tomography system for acquiring 3D images comprising:
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a transport mechanism for moving a biological object relative to a microscope objective to present varying angles of view; at least one optical illumination source for illuminating the entire biological object; a microscope objective lens having a depth of field, the objective lens being located to receive light emanating through the object; an axial translation mechanism coupled to translate the microscope objective lens for axially scanning the entire biological object with the microscope objective lens at a fixed viewpoint through the thickness of the biological object where an axial scan is performed at each of said varying angles of view; at least one ultraviolet camera for sensing a first radiation bandwidth transmitted through the entire biological object and the microscope objective lens during axially scanning, wherein the first radiation bandwidth has a first spectral range between 150 nm and 390 nm to produce a first plurality of pseudo-projection images of the specimen onto the ultraviolet camera, and for sensing a second radiation bandwidth within a second spectral range which is also transmitted through the entire biological object and the microscope objective to the ultraviolet camera during axially scanning, wherein the second spectral range is also between 150 nm and 390 nm, but different than the first spectral range, to produce a second plurality of pseudo-projection images of the specimen onto the ultraviolet camera; wherein each axial scan is performed during a single exposure of the ultraviolet camera, and wherein each pseudo-projection image thus formed includes an integration of a range of focal plane images; an image processor coupled to receive data from the at least one ultraviolet camera; and a reconstruction module coupled to the image processor for reconstructing the plurality of pseudoprojection images to form a reconstructed 3D image. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63)
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Specification