A method of
in situ analysis of a biological sample comprising the steps of (a)
staining the biological sample with N stains of which a first
stain is selected from the group consisting of a first immunohistochemical
stain, a first histological
stain and a first
DNA ploidy stain, and a second stain is selected from the group consisting of a second immunohistochemical stain, a second histological stain and a second
DNA ploidy stain, with provisions that N is an integer greater than three and further that (i) if the first stain is the first immunohistochemical stain then the second stain is either the second histological stain or the second
DNA ploidy stain; (ii) if the first stain is the first histological stain then the second stain is either the second immunohistochemical stain or the second DNA ploidy stain; whereas (iii) if the first stain is the first DNA ploidy stain then the second stain is either the second immunohistochemical stain or the second histological stain; and (b) using a
spectral data collection device for collecting
spectral data from the biological sample, the
spectral data collection device and the N stains are selected such that a
spectral component associated with each of the N stains is collectable.