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Systems and methods for correcting error in biological response signal data

  • US 20050130215A1
  • Filed: 01/24/2005
  • Published: 06/16/2005
  • Est. Priority Date: 12/28/1998
  • Status: Abandoned Application
First Claim
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1. A method of fluorophore bias removal comprising the steps of:

  • (a) labeling a first pool of genetic matter, derived from a biological system representing a baseline state, with a first fluorophore to obtain a first pool of fluorophore-labeled genetic matter;

    (b) labeling a second pool of genetic matter, derived from a biological system representing a perturbed state, with a second fluorophore to obtain a second pool of fluorophore-labeled genetic matter;

    (c) labeling a third pool of genetic matter, derived from said biological system representing said baseline state, with said second fluorophore to obtain a third pool of fluorophore-labeled genetic matter;

    (d) labeling a fourth pool of genetic matter, derived from said biological system representing said perturbed state, with said first fluorophore to obtain a fourth pool of fluorophore-labeled genetic matter;

    (e) contacting said first pool of fluorophore-labeled genetic matter and said second pool of fluorophore-labeled genetic matter with a first microarray under conditions such that hybridization can occur, respectively detecting a first and second flourescent emission signal at each of a plurality of discrete loci on the first microarray from said first and second pool of fluorophore-labeled genetic matter that is bound to said first microarray under said conditions, and determining a first color ratio between said first flourescent emission signal and said second flourescent emission signal;

    (f) contacting said third pool of fluorophore-labeled genetic matter and said fourth pool of fluorophore-labeled genetic matter with a second microarray under conditions such that hybridization can occur, respectively detecting a third and fourth flourescent emission signal at each of a plurality of discrete loci on the second microarray from said third and fourth pool of fluorophore-labeled genetic matter that is bound to said second microarray under said conditions, and determining a second color ratio between said third flourescent emission signal and said fourth flourescent emission signal; and

    (g) computing an average color ratio by averaging said first color ratio and said second color ratio;

    wherein;

    said first microarray and said second microarray are similar to each other, exact replicas of each other, or are identical; and

    said first color ratio and said second color ratio are determined for the same individual component of genetic matter, thereby removing fluorophore bias.

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