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Optical Projection Tomography for 3D Microscopy Disclosure Number: IPCOM000030403D
Publication Date: 2004-Aug-11
Document File: 13 page(s) / 39K

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An optical image reconstructing apparatus comprising a) a laser; b) an optical means for focusing a laser beam from said laser within the volume of an sample object, collecting light emitted, and directing the light collected toward one or more photosensitive detectors; c) an image pickup means for converting the images of said object into respective electrical image signals, and for outputting said respective electrical image signals; d) a moving means for successively rotating said object at a predetermined angular interval about an axis which intersects a point on an optical axis of said first optical means, and translating said sample in a direction perpendicular to said rotation axis; e) software means for deconvoluting electronic signals received from said photosensitive detectors into three-dimensional image; and f) means for displaying said three-dimensional image; whereby the three dimensional fine structure of said sample can be easily visualized.

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Optical Projection Tomography for 3D Microscopy

Field of the Invention

This invention concerns an optical image reconstruction apparatus and method. More specifically, the invention relates to an optical projection tomography apparatus, particularly for use in fluorescence imaging.

Background of the Invention

In one form of X-ray computed tomography, the subject is aligned relative to the source in a given angular direction. The source shoots parallel, collimated beams of X-rays cutting across the subject perpendicular to the subject’s axis and a detector collects the attenuated X-rays on the other side of the subject. The source and detector are then turned a small angular amount relative to the subject and another X-ray image is gathered. This continues until the images are gathered from all angular directions so a reasonable two-dimensional image of the subject can be reconstructed. The source and detector are then repositioned at another point along the subject’s axis and another set of images are gathered. This process is continued depending on the region of interest in the subject.

WO0259562 describes an optical coherence microscope. The scanner described by US patent 5,680,484 uses conventional light microscopy to gather image information. The patent does not describe an excitation source but it is assumed to be ambient light. Instead of projecting parallel cones of light into the subject, it simply gathers light from a plane within the subject where the lens is focused. After this image is gathered, the lens is refocused to a different plane within the subject and another image is gathered. After multiple images are gathered from multiple focal planes for one angular orientation, all the images are added together to simulate one image formed from one X-ray projection. This process of gathering light from multiple focal planes is repeated for multiple angles. US patent 5,680,484.

Brief Description of the Figures

Figure 1 is an optical diagram of one embodiment of the current invention.

Figure 2 shows the optics of one embodiment of the current invention.

Figure 3 shows the instrument platform of one embodiment of the current invention.

Figures 4A, 4B and 4C are diagrams of a sample chamber and associated stage for rotational and linear translation of the sample.

Figures 5A and 5B shows one example of a sample chamber and associated stage for rotational and linear translation of the sample.

Detailed Description of the Invention

The OPT system in the current invention uses visible light as an excitation source and the subject is moved relative to the source and detector. By using epi-fluorescent optics, the scanner projects a cone of light several millimeters into the subject and then gathers emitted fluorescent light from the excited volume. The geometry of the excitation cone makes it appear as almost a cylinder of light. This excitation cone penetrates roughly half way into the subject. This cone of light is projected into the sub...