Systems and methods for imaging and processing tissue
Inventors
Assignees
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Publication Number
US-9983134-B2
Publication Date
2018-05-29
Expiration Date
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, capturing the sectioned 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, and capturing each sectioned tissue, 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. Further, captured tissue sections can be processed, re-imaged, and indexed to their original location in the three dimensional image.
Core Innovation
The invention provides a method of imaging tissue that illuminates a first region of the tissue with light and detects light from the tissue in response to the illuminating light to form at least a first image of the first region. The detected light includes light received from a depth in the tissue below a first sectioning depth to provide first registration data. A portion of the tissue is sectioned at the first sectioning depth to expose a second region, and the second region is illuminated and imaged.
The method processes the images of the first region and the second region including at least a first optical tomography image and a second optical tomography image. The images are indexed using the first registration data and the second registration data to form a three dimensional image that includes at least the first optical tomography image and the second optical tomography image. In this way, optical tomography images from multiple tissue regions are combined into a three dimensional image by indexing based on registration data derived from depths below respective sectioning depths.
For brain tissue, the invention further defines a method of imaging brain tissue in which a first region is illuminated and light is detected to form at least a first image. The method then sectiones a portion of the brain tissue to expose a second region using a sectioning tool, positions the exposed second region relative to a scanning lens using a translation stage connected to an automated control system, and illuminates and detects light from the second region to form at least a second image. The images of the first and second regions are processed with a data processor to include at least a first optical tomography image and a second optical tomography image.
Claims Coverage
The document contains two independent claims. Together, they define a multi-stage imaging workflow with optical tomography and indexing or processing to form three dimensional imaging output, grounded in registration data derived from detected light below sectioning depths.
Optical tomography imaging across sectioned regions with registration data from below sectioning depths
Illuminating a first region of the tissue with light; detecting light to form a first image including light received from a depth below a first sectioning depth to provide first registration data; sectioning at the first sectioning depth to expose a second region; illuminating the second region and detecting light to form a second image including light received from a depth below a second sectioning depth to provide second registration data; processing the images including at least a first optical tomography image and a second optical tomography image; and indexing the images using the first and second registration data to form a three dimensional image including at least the first optical tomography image and the second optical tomography image.
Brain tissue imaging with translation stage positioning and processing into optical tomography images
Illuminating a first region of the brain tissue with light; detecting light to form at least a first image; sectioning a portion to expose a second region with a sectioning tool; positioning the exposed second region relative to a scanning lens using a translation stage connected to an automated control system; illuminating the second region and detecting light to form at least a second image; and processing the images with a data processor to include at least a first optical tomography image of the first region and a second optical tomography image of the second region.
Across both independent claims, the core inventive concept is imaging multiple regions that result from sectioning, using detected light from depths below respective sectioning depths to provide registration data and producing optical tomography images that are combined into three dimensional imaging output through indexing or processing.
Stated Advantages
Reduced registration artifacts vs. block-face approaches.
Preserved samples for downstream biochemical assays including immunohistochemistry (IHC), FISH, mass spectrometry, and re-staining.
Automation.
Documented Applications
Whole mouse heart imaging with multispectral detection, including reimaging/overlay back onto original volumes and registration for downstream analyses (e.g., angiogenesis assay correlations using IHC/EF5 hypoxia, collagen SHG, nuclei, and vascular cast PU4ii).
Automated generation of mouse brain atlases from captured and registered section data.
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