System, method, and kit for processing a magnified image of biological material to identify components of a biological object
Inventors
Hunter, Edward • Mc Donough, Patrick M. • Mikic, Ivana • Price, Jeffrey H.
Assignees
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Publication Number
US-7933435-B2
Publication Date
2011-04-26
Expiration Date
Abstract
A system, method and kit for processing an original image of biological material to identify certain components of a biological object by locating the biological object in the image, enhancing the image by sharpening components of interest in the object, and applying a contour-finding function to the enhanced image to create a contour mask. The contour mask may be processed to yield a segmented image divided by structural units of the biological material.
Core Innovation
The invention relates to a computerized method for analyzing an original magnified image of cells using a high throughput microscopy system. The method generates membrane masks by enhancing traces of membrane outlines in a normalized original image using at least a first enhancement function and thresholding the enhanced traces.
A first enhanced membrane image is generated from the first membrane mask, and a second membrane mask is generated using at least a second enhancement function followed by thresholding to produce a second enhanced membrane image. The method generates a composite enhanced membrane image by combining the first and second enhanced membrane images.
It then generates markers associated with nuclei of the cells and generates a contour mask of membrane outlines with a contour finding function that applies the markers to the composite enhanced membrane image. An enhanced membrane mask is generated from the contour mask, and structures of cellular components are measured in the original image using the enhanced membrane mask.
In embodiments described, contour finding uses a reconstruction algorithm including watershed contour mask reconstruction applied with markers associated with nuclei or lipid markers for lipid droplets. The system also includes options for reconstructing masks by thickening contour lines to preserve structure, followed by compartment segmentation and measurement outputs such as cell counts, dimensions, areas, intensities, and ratios.
Claims Coverage
The independent claim covers a computerized high-throughput microscopy workflow with multiple membrane-outline enhancement functions, composite enhanced membrane image generation, nucleus-associated marker-driven contour finding, and measurement of cellular component structures using an enhanced membrane mask. Dependent claims refine the contour finding and enhanced membrane mask generation steps.
Generating a first and second membrane masks using multiple enhancement functions and thresholding
Generating a first membrane mask by enhancing the traces of membrane outlines in cells in a normalized original image using at least a first enhancement function and then thresholding the enhanced traces; generating a first enhanced membrane image from the first membrane mask; generating a second membrane mask by enhancing the traces of membrane outlines in cells in the normalized original image using at least a second enhancement function and then thresholding the enhanced traces; generating a second enhanced membrane image from the second membrane mask.
Creating a composite enhanced membrane image from multiple enhanced membrane images
Generating a composite enhanced membrane image by combining the first and second enhanced membrane images.
Marker-assisted contour mask generation using nucleus-associated markers
Generating markers associated with nuclei of the cells; generating a contour mask of membrane outlines with a contour finding function that applies the markers to the composite enhanced membrane image.
Generating an enhanced membrane mask from the contour mask and measuring cellular components
Generating an enhanced membrane mask from the contour mask; measuring structures of cellular components in the original image with the enhanced membrane mask.
Watershed-based contour finding
The contour finding function that applies the markers to the composite enhanced membrane image uses a watershed algorithm.
Thickening contour lines to generate the enhanced membrane mask
Generating an enhanced membrane mask by thickening the lines of the contour mask.
Overall, the claim coverage centers on producing an enhanced membrane mask by combining composite enhanced membrane imaging from multiple enhancement functions with a nucleus-marker-applied contour finding function, optionally using watershed and thickening to refine the resulting membrane-outline mask for downstream measurement of cellular component structures.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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