Apparatus and method for imaging small objects in a flow stream using optical tomography
First Claim
1. A flow optical tomography method for imaging and analysis of microscopic objects, the method comprising the steps of:
- (a) injecting at least one object into an injection tube;
(b) controlling the flow of the at least one object through a capillary tube such that the at least one object elongates along an axis of flow and moves proximately along a central axis of the capillary tube;
(c) sampling with at least one optical point source, located around a circumference of the capillary tube, in conjunction with at least one opposing optical sensor disposed opposite the at least one optical point source at a distance from the capillary tube such that there is no focal plane within the at least one object, and where multiple projection angles through the at least one object are sampled as it flows past the at least one optical point source and at least one opposing optical sensor; and
(d) generating a series of timing signals, such that each timing signal coincides with a specific position along the axis in the z-direction of the at least one object, so as to generate a set of timed optical projections through the at least one object.
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Abstract
A flow optical tomography system includes a flow cytometer, and at least one reconstruction cylinder positioned around a capillary tube. A photon source and a photon sensor work together with a pulse height analyzer to provide a first trigger point for the beginning of an object or cell, and a second trigger point for the end of the cell. The trigger signal is received by the reconstruction cylinder. The reconstruction cylinder includes optical point sources having a selectable emission wavelength, disposed in a geometric pattern around the cylinder perpendicular to and concentric with the capillary tube axis that facilitate the acquisition of transmitted, attenuated projection images of the flowing cells. The sensors also collect projections of fluorescence emitted from tagged molecular probes associated with nuclear and/or cytoplasmic structures or cell membranes. The projections are algorithmically processed to provide three dimensional information about the cells and their disease state.
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Citations
66 Claims
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1. A flow optical tomography method for imaging and analysis of microscopic objects, the method comprising the steps of:
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(a) injecting at least one object into an injection tube;
(b) controlling the flow of the at least one object through a capillary tube such that the at least one object elongates along an axis of flow and moves proximately along a central axis of the capillary tube;
(c) sampling with at least one optical point source, located around a circumference of the capillary tube, in conjunction with at least one opposing optical sensor disposed opposite the at least one optical point source at a distance from the capillary tube such that there is no focal plane within the at least one object, and where multiple projection angles through the at least one object are sampled as it flows past the at least one optical point source and at least one opposing optical sensor; and
(d) generating a series of timing signals, such that each timing signal coincides with a specific position along the axis in the z-direction of the at least one object, so as to generate a set of timed optical projections through the at least one object. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 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, 60, 61, 62, 63, 64, 65, 66)
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17. A flow optical tomography system for imaging and analysis of microscopic objects, comprising:
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(a) a flow cytometer including a capillary tube;
(b) at least one reconstruction cylinder positioned around the capillary tube; and
(c) a triggering device, located to view the at least one object flowing through the capillary tube, for creating a trigger signal for at least one object, where the trigger signal is received by the at least one reconstruction cylinder, and where the at least one reconstruction cylinder responds to the trigger signal by producing signals representing a projection image about the at least one object and the signals are processed to provide three dimensional information about the at least one object.
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59. A flow optical tomography system for imaging and analysis of microscopic objects, the method comprising:
- a pulse height analyzer, a source of photons and a photon sensor, where the source of photons and the photon sensor work together with the pulse height analyzer to operate as a triggering device, where the pulse height analyzer provides a first trigger point for the beginning of a the at least one object, and a second trigger point for the end of the at least one object to create a corresponding trigger signal delivered to a reconstruction cylinder for the purpose of synchronizing object velocity and position with each projection slice.
Specification