A system for determining a score representative of a proximity between at least one pair of cells in a tumor tissue sample, the system comprising: a programmable controller programmed to: receive image data from an imaging device, separate the image data into unmixed image data, and provide the data through a plurality of data channels; wherein the unmixed image data in a first data channel describes fluorescence signals attributable to a cell nucleus, the unmixed data in a second data channel describes fluorescence signals attributable to tumor tissue, the unmixed data in a third data channel describes fluorescence signals attributable to a first biomarker, and the unmixed image data in a fourth data channel describes fluorescence signals attributable to a second biomarker; receive image data from the first data channel and generating an image representative of all nuclei in a field of view and dilate the image to generate a cell mask; receive image data from the second data channel and generate a mask of all tumor area in the field of view; receive image data from the third data channel and generate a first biomarker mask of all cells in the field of view that express the first biomarker; receive image data from the fourth data channel and generate a second biomarker mask of all cells in the field of view that express the second biomarker; combine the cell mask and the tumor area mask to generate at least one of a non-tumor cell mask and a tumor cell mask; dilate the first biomarker mask by a margin sufficient to encompass proximally located cells expressing the second biomarker to generate a dilated first biomarker mask; combine the dilated first biomarker mask and the second biomarker mask to generate an interaction mask; calculate a spatial proximity score based on the area of the interaction mask and the area of at least one of the non-tumor cell mask, tumor cell mask, and cell mask. receive image data from an imaging device, separate the image data into unmixed image data, and provide the data through a plurality of data channels; wherein the unmixed image data in a first data channel describes fluorescence signals attributable to a cell nucleus, the unmixed data in a second data channel describes fluorescence signals attributable to tumor tissue, the unmixed data in a third data channel describes fluorescence signals attributable to a first biomarker, and the unmixed image data in a fourth data channel describes fluorescence signals attributable to a second biomarker; receive image data from the first data channel and generating an image representative of all nuclei in a field of view and dilate the image to generate a cell mask; receive image data from the second data channel and generate a mask of all tumor area in the field of view; receive image data from the third data channel and generate a first biomarker mask of all cells in the field of view that express the first biomarker; receive image data from the fourth data channel and generate a second biomarker mask of all cells in the field of view that express the second biomarker; combine the cell mask and the tumor area mask to generate at least one of a non-tumor cell mask and a tumor cell mask; dilate the first biomarker mask by a margin sufficient to encompass proximally located cells expressing the second biomarker to generate a dilated first biomarker mask; combine the dilated first biomarker mask and the second biomarker mask to generate an interaction mask; calculate a spatial proximity score based on the area of the interaction mask and the area of at least one of the non-tumor cell mask, tumor cell mask, and cell mask.