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Brendon J Coventry1, Michael J Weightman1, John M Skinner2, John Bradley31Discipline of Surgery, University of Adelaide; 2Department of Pathology; 3Department of Clinical Immunology, Flinders University, Adelaide, SA, AustraliaAbstract: Quantitation of cell density in tissues has proven problematic over the years. The manual microscopic methodology, where an investigator visually samples multiple areas within slides of tissue sections, has long remained the basic 'standard' for many studies and for routine histopathologic reporting. Nevertheless, novel techniques that may provide a more standardized approach to quantitation of cells in tissue sections have been made possible by computerized video image analysis methods over recent years. The present study describes a novel, computer-assisted video image analysis method of quantitating immunostained cells within tissue sections, providing continuous graphical data. This technique enables the measurement of both distribution and density of cells within tissue sections. Specifically, the study considered immunoperoxidase-stained tumor infiltrating lymphocytes within breast tumor specimens, using the number of immunostained pixels within tissue sections to determine cellular density and number. Comparison was made between standard manual graded quantitation methods and video image analysis, using the same tissue sections. The study demonstrates that video image techniques and computer analysis can provide continuous data on cell density and number in immunostained tissue sections, which compares favorably with standard visual quantitation methods, and may offer an alternative.Keywords: immunostaining, video image analysis, cellular quantitation, tissue sections, breast cancer, tumor infiltrating lymphocytes