Patent ID: 8639013

Claim:
A method for generating a brightfield type image that resembles a brightfield staining protocol comprising the steps of: acquiring image data of two or more fluorescent images of a fixed area on a biological sample; analyzing the image data utilizing, at least in part, featured based information or pixel intensity data information to generate mapping parameters wherein said mapping parameters comprises a nonlinear estimation model; applying said mapping parameters to the fluorescent images; transforming the two or more fluorescent imaging into a brightfield color space; and generating a brightfield type image. where the nonlinear estimation model is defined as: R= 255 exp(− a[ Dye1]* z[ Dye1]− a[ Dye2] z[ Dye2]−. . . a[ Dyen] z[ Dyen]) G= 255 exp(− b[ Dye1]* z[ Dye1]− b[ Dye2] z[ Dye2]−. . . ) B =255 exp(− c[ Dye1]* z[ Dye1]− c[ Dye2] z[ Dye2]−. . . ) wherein R, G, and B are resulting red, green and blue pixel values in the brightfield type image; z is a scaling coefficient for a fluorescent dye quantities observed at a given pixel location; and a, b, and c are the extinction coefficients corresponding to the brightfield color space, and wherein the triples, a[Dyen], b[Dyen], c[Dyen], are a constant times the extinction coefficients of the nth dye in the virtual stain as defined using a preselected or desired color.