Patent ID: 6456734
Filing Date: 2002-09-24
Classification: G01N

Abstract:
A method for transforming an initial FRET image of a specimen to a processed FRET image, the initial image being represented by a set of pixels each having three color space coordinates, the method including the steps of:(a) generating a corrected FRET image from the initial image by correcting for spectral overlap by applying a transformation of the following form to obtain a corrected FRET channel image Fc: Fc=Fbâˆ’(Fdb/Ddb)Ã—Dbâˆ’(Fab/Aab)Ã—Ab wherein Fb are pixels of an initial FRET channel image, Db are pixels of an initial donor channel image, Ab are pixels of an initial acceptor channel image, Fb are donor pixels of the initial FRET channel image, Fab are acceptor pixels of the initial FRET channel image, Ddb are donor pixels of the initial donor channel image, and Aab are acceptor pixels of the initial acceptor channel image, all after background subtraction;(b) selecting at least two image regions in the corrected FRET image, each image region including at least one pixel, wherein the image regions define at least two distinguishable spectral categories and do not substantially overlap spatially; (c) performing a color space transformation on the corrected image, based on the color space coordinates of the pixels in each of the selected image regions, to generate a processed image; and (d) evaluating the processed image for a visible indication of an improved spatial distribution of color.