System and method for automatic color segmentation and minimum significant response for measurement of fractional localized intensity of cellular compartments
Inventors
Hunter, Edward A. • Ingermanson, Randall Scott • Callaway, William Scott
Assignees
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Publication Number
US-8116982-B2
Publication Date
2012-02-14
Expiration Date
Abstract
A system, a method, and a programmed device for measurement of translocational activity among cellular compartments process magnified images of cellular material exposed to an agent by segmenting and compartmentalizing the images and then measuring fractional localized intensity of two or more components in the segmented, compartmentalized image. The measured fractional localized intensities are compared to determine translocation of cellular material among the measured components caused by the agent.
Core Innovation
The invention relates to a method executable by an automated microscopy platform for measuring cell activity represented in an image of cells treated with an agent. The method comprises segmenting cellular components in the image, separating segmented components of overlapping cells in the image, and determining translocation of cellular material between first cellular compartments and second cellular compartments caused by the agent in the segmented, separated image.
Translocation is determined by measuring a first intensity of the cellular material in a first cellular compartment and a second intensity of the cellular material in a second cellular compartment of the cell. Fractional localized intensity F_a of the cellular material in the first cellular compartment is determined according to a ratio of the first intensity to a sum of at least the first and second intensities, based on summed pixel emission intensities across compartments.
The emission intensity I_i(x,y) at a pixel location (x,y) is determined according to a formulation that includes an incident excitation intensity I_0, a quantum yield Q, an extinction coefficient ε, a local fluorophore concentration u at the pixel location, and a thickness z of a column of cellular material at the pixel location. The disclosed dependences refine segmenting by enhancing contrast between cellular compartments and the image background and refine separating by tessellating the image.
Claims Coverage
The document provides one independent claim, directed to an automated microscopy method that combines segmentation and separation of overlapping cells with compartment-specific fractional localized intensity to determine agent-induced translocation. Dependent claims narrow the contrast basis for segmentation and specify tessellating-based separation, with one dependency combining both refinements.
Automated microscopy segmentation and overlap separation for agent response
A method executable by an automated microscopy platform for measuring cell activity represented in an image of cells treated with an agent, where the method comprises segmenting cellular components; separating segmented components of overlapping cells; and determining, in the segmented, separated image, translocation of cellular material between first cellular compartments and second cellular compartments caused by the agent.
Fractional localized intensity based on compartment intensity ratio
Determining fractional localized intensity F_a of the cellular material in the first cellular compartment according to a ratio of the first intensity to a sum of at least the first and second intensities, based on summed pixel emission intensities in compartment a relative to compartments a and b.
Emission intensity model for fractional localized intensity
Determining emission intensity I_i(x,y) at a pixel location (x,y) according to I_i(x,y)=∫_0^z I_0·Q·ε·u·dz, where I_0 is an incident excitation intensity, Q is a quantum yield, ε is an extinction coefficient, u is a local fluorophore concentration at pixel location (x,y), and z is the thickness of a column of cellular material at pixel location (x,y).
Contrast-enhanced segmentation relative to image background
Segmenting further comprises enhancing contrast between the first cellular compartments and the image background, and between the second cellular compartments and the image background.
Tessellating-based separation of overlapping cells
Separating further includes tessellating the image.
Combined contrast-enhanced segmentation and tessellating separation
Segmentation is performed by enhancing contrast between cellular compartments and the image background, and separation is performed by tessellating the image.
Across the independent claim and its dependencies, the claims focus on automated microscopy image processing and determining agent-induced translocation by computing fractional localized intensity from compartment-specific emission intensities using a defined emission intensity model; dependent claims further specify contrast-enhanced segmentation and tessellation-based separation.
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