A method of determining an amount of at least one molecular species in a
sample from an image of the sample captured by an image acquisition
device is provided, each molecular species being indicated by a dye. A
dye space representation of a plurality of dyes is formed by orthogonally
adding the correspondence tables of the dyes, each correspondence table
having a plurality of normalized RGB triplets and incrementally extending
from 0% to 100% transmittance. The dye space representation has one
dimension for each dye and provides a reference model for a combination
of the plurality of dyes. Each pixel of an image of the sample stained
with the combination of the plurality of dyes is compared to the
reference model, each pixel having a color defined by an RGB triplet, so
as to determine an optimal combination of normalized RGB triplets from
the respective correspondence tables of the dyes producing the color of
the respective pixel. An artificial image of the sample is then formed
from the normalized RGB triplets for each dye as determined from the
optimal combination. The artificial image thereby indicates a
distribution of the respective dye over the sample image and facilitates
determination of the amount of the corresponding molecular species.
Associated methods, systems, and computer software program products are
also provided.