Cluster analysis of genetic microarray images
First Claim
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1. A method of determining the relative abundance of genetic material present in a first sample and a second sample, comprising the steps of:
- labeling the first sample with a first detectable label adapted to bind with genetic material therein;
labeling the second sample with a second detectable label adapted to bind with genetic material therein;
providing a substrate with a binding area thereon for hybridizing with a genetic sequence;
contacting the substrate comprising said genetic sequence with the first and second samples under conditions suitable to effect hybridization;
obtaining a digital image of the binding area under conditions causing said labels to manifest an image intensity dependent on the abundance of genetic material bound to the binding area, if any, thus providing an array of pixel values comprising respective intensity values corresponding the first and second labels;
assigning each of said pixel values to a respective one of a foreground cluster and a background cluster;
determining a representative statistical vector of pixel values within each of said foreground cluster and said background cluster; and
computing the relative abundance of genetic material within the first and second samples corresponding to the binding area in proportion to a ratio of differences between the respective coordinates of the representative statistical vectors.
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Abstract
A method for determining relative incidence of a binding substance within two biological samples is provided. The two samples are labeled with luminescent materials having different chromatic properties. An image of the luminescent materials upon a binding site of a microarray is analyzed as two clusters of data points scattered about respective representative pairs of chromatic intensity values. The relative incidence of the binding substance is determined as a ratio of differences between corresponding indices of the representative pairs.
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Citations
6 Claims
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1. A method of determining the relative abundance of genetic material present in a first sample and a second sample, comprising the steps of:
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labeling the first sample with a first detectable label adapted to bind with genetic material therein; labeling the second sample with a second detectable label adapted to bind with genetic material therein; providing a substrate with a binding area thereon for hybridizing with a genetic sequence; contacting the substrate comprising said genetic sequence with the first and second samples under conditions suitable to effect hybridization; obtaining a digital image of the binding area under conditions causing said labels to manifest an image intensity dependent on the abundance of genetic material bound to the binding area, if any, thus providing an array of pixel values comprising respective intensity values corresponding the first and second labels; assigning each of said pixel values to a respective one of a foreground cluster and a background cluster; determining a representative statistical vector of pixel values within each of said foreground cluster and said background cluster; and computing the relative abundance of genetic material within the first and second samples corresponding to the binding area in proportion to a ratio of differences between the respective coordinates of the representative statistical vectors. - View Dependent Claims (2, 3, 4, 5, 6)
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Specification