Phenotyping and quantitation of immune cells using multi-parameter imaging of solid tissue is likely to be essential for evaluating the effects and effectiveness of novel immunotherapy. Multi-parameter flow cytometry is commonly used for phenotyping and quantitating cells in peripheral blood, and is well understood in clinical settings, but cannot be used to quantitate cells in solid tissues. Analysis of tissue pathology remains largely restricted to measuring histological parameters (size of pathology) or single colours (e.g Dab staining). We have therefore been evaluating the use of flow cytometry software for analysing multiparameter images of histology slides stained with fluorescent dyes. We have developed strategies to open, merge images, and merge cell segmentation data in flow cytometry software. The data generated from multiple regions of interest by fluorescent slide scanners such as the Vectra Pathology microscope can now be easily merged to generate large numbers of events for analysis. The data can be visualised or mined using standard gating or more complex algorithms such as tSNE, and then back gated to the image to locate or verify any cells of interest. The images can also be used as part of the gating strategy to segment the tissue for analysis. The benefits of this strategy are that spectral un-mixing can be used to accurately generate and analyse complex multi-parameter data in different tissue areas. The data can then be analysed in a way that is easily understood by immunology researchers and clinicians who are familiar with results produced from flow cytometry software.