Quantitative assessment of biological function based on the temporal and spatial organization of subnuclear domains
First Claim
1. A method for quantifying domains within a punctate distribution comprising the steps of:
- acquiring at least one digital image of a punctate distribution, wherein the at least one digital image is a fluorescence photomicrographic image of the nucleus of a eukaryotic cell;
performing image segmentation on the at least one digital image to produce a segmented image of the punctate distribution;
identifying domains within the punctate distribution of the segmented image; and
computing at least twenty-five organizational parameters of the domains within the punctate distribution of the segmented image wherein the at least 25 organizational parameters comprise the number of domains, mean domain size, median domain size, standard deviation of domain size, variance of domain size, skewness of domain size, kurtosis of domain size, coefficient of variation of domain size;
index of dispersion of domain size;
mean nearest neighbor distance, median nearest neighbor distance, standard deviation of nearest neighbor distance, variance of nearest neighbor distance, skewness of nearest neighbor distance, kurtosis of nearest neighbor distance, coefficient of variation of nearest neighbor distance;
index of dispersion of nearest neighbor distance, domain density, Re, Ro/Re, Ve, Vo/Ve, Rp, Rd, or Rd/Rp.
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Abstract
The present invention provides a method useful for analysis of punctate distribution of supramolecular complexes that are compartmentalized in nuclear microenvironments. In preferred embodiments the method provides quantitative analysis of the assembly of regulatory machinery within the context of nuclear architecture and to provide a mechanistic link with biological control. In other embodiments, the invention provides a method of identifying a molecule by parameters of its subnuclear distribution, comprising the steps of quantifying organizational parameters of domains within subnuclear distributions of a molecule; performing factor analysis on the quantified organizational parameters; performing discriminant analysis on the quantified organizational parameters; comparing the results of the factor analysis and the results of the discriminant analysis to one or more standards, thereby identifying a molecule by parameters of its subnuclear distribution.
9 Citations
23 Claims
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1. A method for quantifying domains within a punctate distribution comprising the steps of:
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acquiring at least one digital image of a punctate distribution, wherein the at least one digital image is a fluorescence photomicrographic image of the nucleus of a eukaryotic cell; performing image segmentation on the at least one digital image to produce a segmented image of the punctate distribution; identifying domains within the punctate distribution of the segmented image; and computing at least twenty-five organizational parameters of the domains within the punctate distribution of the segmented image wherein the at least 25 organizational parameters comprise the number of domains, mean domain size, median domain size, standard deviation of domain size, variance of domain size, skewness of domain size, kurtosis of domain size, coefficient of variation of domain size;
index of dispersion of domain size;
mean nearest neighbor distance, median nearest neighbor distance, standard deviation of nearest neighbor distance, variance of nearest neighbor distance, skewness of nearest neighbor distance, kurtosis of nearest neighbor distance, coefficient of variation of nearest neighbor distance;
index of dispersion of nearest neighbor distance, domain density, Re, Ro/Re, Ve, Vo/Ve, Rp, Rd, or Rd/Rp. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11)
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12. A method for quantifying domains within a punctate distribution comprising the steps of:
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acquiring at least one digital image of a punctate distribution, wherein the at least one digital image is a fluorescence photomicrographic image of the nucleus of a eukaryotic cell; performing image segmentation on the at least one digital image to produce a segmented image of the punctate distribution; identifying protein domains within the punctate distribution in the segmented image; computing organizational parameters of the protein domains within the punctate distribution of the segmented image; and performing statistical analysis of the organizational parameters wherein the organizational parameters comprise the number of domains, mean domain size, median domain size, standard deviation of domain size, variance of domain size, skewness of domain size, kurtosis of domain size, coefficient of variation of domain size;
index of dispersion of domain size;
mean nearest neighbor distance, median nearest neighbor distance, standard deviation of nearest neighbor distance, variance of nearest neighbor distance, skewness of nearest neighbor distance, kurtosis of nearest neighbor distance, coefficient of variation of nearest neighbor distance;
index of dispersion of nearest neighbor distance, domain density, Re, Ro/Re, Ve, Vo/Ve, Rp, Rd, or Rd/Rp. - View Dependent Claims (13, 14, 15, 16, 17, 18)
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19. A method for quantifying the organization of molecules in a cell, comprising:
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acquiring a digital image that comprises fluorescently labeled molecules in a cell, wherein the digital image is a fluorescence photomicrographic image; segmenting the at least one digital image to produce a segmented image; detecting, in the segmented image, domains of the molecules in the cell; determining the number of domains in the cell; determining the areas of the domains and the centroids of the domains; determining the locations of the centroids of the domains in pixel coordinates of the segmented image wherein the at least 25 organizational parameters comprise the number of domains, mean domain size, median domain size, standard deviation of domain size, variance of domain size, skewness of domain size, kurtosis of domain size, coefficient of variation of domain size;
index of dispersion of domain size;
mean nearest neighbor distance, median nearest neighbor distance, standard deviation of nearest neighbor distance, variance of nearest neighbor distance, skewness of nearest neighbor distance, kurtosis of nearest neighbor distance, coefficient of variation of nearest neighbor distance;
index of dispersion of nearest neighbor distance, domain density, Re, Ro/Re, Ve, Vo/Ve, Rp, Rd, or Rd/Rp; andcomputing at least twenty-five parameters that quantify the organization of the molecule in the cell based, in part, on the areas and the locations of the centroids of the domains. - View Dependent Claims (20, 21, 22, 23)
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Specification