Systems and methods for volumetric tissue scanning microscopy
First Claim
1. A method for imaging tissue, comprising the steps of:
- mounting the tissue on a computer controlled stage of a microscope;
determining volumetric imaging parameters;
directing at least two photons onto a region of interest;
scanning the region of interest across a portion of the tissue;
imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest;
sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volume of the tissue in the region of interest;
detecting an image of the tissue due to said excitation light; and
processing three-dimensional data that is imaged to create a three-dimensional image of the region of interest.
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Abstract
In accordance with preferred embodiments of the present invention, a method for imaging tissue, for example, includes the steps of mounting the tissue on a computer controlled stage of a microscope, determining volumetric imaging parameters, directing at least two photons into a region of interest, scanning the region of interest across a portion of the tissue, imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest, sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volumes of the tissue in the region of interest, detecting a fluorescence image of the tissue due to said excitation light; and processing three-dimensional data that is collected to create a three-dimensional image of the region of interest.
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Citations
29 Claims
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1. A method for imaging tissue, comprising the steps of:
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mounting the tissue on a computer controlled stage of a microscope;
determining volumetric imaging parameters;
directing at least two photons onto a region of interest;
scanning the region of interest across a portion of the tissue;
imaging a plurality of layers of the tissue in a plurality of volumes of the tissue in the region of interest;
sectioning the portion of the tissue and imaging a second plurality of layers of the tissue in a second plurality of volume of the tissue in the region of interest;
detecting an image of the tissue due to said excitation light; and
processing three-dimensional data that is imaged to create a three-dimensional image of the region of interest. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10)
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11. A system for providing a three-dimensional image of a region of interest, comprising:
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a light source for producing excitation light and providing at least two photons onto the region of interest;
a scanning microscope optically coupled to the light source;
a tissue sectioning device mechanically coupled to the microscope;
an x-y scanner to scan the region of interest;
an image sensor that detects a plurality of images of the region of interest; and
a data processor that processes the plurality of images to produce a processed three-dimensional image of the region of interest. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23)
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24. A method of imaging tissue in-vivo, comprising the steps of:
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mounting the tissue in a multi-photon microscope;
directing at least two photons onto a region of interest;
scanning a plurality of layers of the tissue in the region of interest;
imaging a plurality of layers in the tissue in the region of interest;
detecting a fluorescence image of the region of interest due to said excitation light; and
processing the detected fluorescence image comprising the steps of;
sequentially storing a plurality of portions of three-dimensional image data set;
enhancing the image data set;
registering individual three-dimensional data sets to generate a large three-dimensional data set; and
displaying the three-dimensional data set of the region of interest. - View Dependent Claims (25, 26, 27, 28, 29)
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Specification