Patent ID: 6453060
Filing Date: 2002-09-17
Classification: G01N,G06K

Abstract:
A method for separating contrasts for images of a biological specimen wherein the biological specimen has been prepared with a chromogen comprising the steps of:(a) obtaining an image of the biological specimen at a wavelength of interest wherein the image comprises a plurality of pixels; (b) measuring the zero attenuation grayscale value from clear areas of the image, wherein the zero attenuation grayscale value is GN(0) and the attenuation coefficient &agr;n(i,j) is: Î±n&it;â€ƒ&it;(i,j)=-ln&it;â€ƒ&it;(Gn&it;â€ƒ&it;(i,j)Gn&it;â€ƒ&it;(0))â€ƒwhere n=index identifying filter with center wavelength &lgr;n; Gn(i,j)=grayscale value for pixel i,j in image collected at &lgr;n; Gn(0)=grayscale value for zero attenuation in specimen using filter n; and &agr;n(i,j)=attenuation coefficient for pixel i,j; (c) computing the attenuation coefficient for all pixels in the image from the zero attenuation grayscale value and pixel values; (d) expressing the attenuation coefficient as the concentration times an extinction coefficient for the chromogen at the wavelength of interest; and (e) solving for the concentration of the chromogen at each pixel by dividing the attenuation by the extinction coefficient.