Method for increasing clinical specificity when detecting tumors and their precursor stages by simultaneously measuring at least two different molecular markers

An automatable method for identifying cancer cells and their precursor cells in a cell sample or tissue sample, said method comprising the following steps:

• a) selecting at least two molecular markers of cancer, wherein the detection of each of said markers alone is not a reliable indicator of the presence of cancer cells and their precursor cells in said cell sample or tissue sample,
• b) contacting the cell sample or tissue sample with signaling reagents that specifically bind to said at least two molecular markers,
• c) simultaneously detecting signal intensities from the markers within a single cell of said cell sample or within a constituent region of a section of said tissue sample,
• d) combining and accrediting the signal intensities detected, and comparing the combined and accredited signal intensities to a threshold value, wherein combined and accredited signal intensities above or below the threshold value indicate the presence of cancer cells and their precursors in said cell sample or tissue sample, wherein steps b)-d) collectively comprise:
• i) staining each marker an individual color that is different from a color that every other marker is stained;
• ii) defining a threshold value for a plurality of secondary colors of mixtures of said individual colors;
• iii) determining the presence of multiple markers with a single cell of said cell sample or within a constituent region of a section of said tissue sample by detecting the secondary color of a mixture of said individual colors within said cell or within said constituent region of said tissue sample section;
• and
• iv) relating the secondary color detected to the threshold value, wherein a value of the secondary color detected above or below the threshold value is predetermined to be indicative of the presence of cancer cells and their precursors in the cell sample or tissue sample.