Abstract
BackgroundCell motility is a critical parameter in many physiological as well as pathophysiological processes. In time-lapse video microscopy, manual cell tracking remains the most common method of analyzing migratory behavior of cell populations. In addition to being labor-intensive, this method is susceptible to user-dependent errors regarding the selection of "representative" subsets of cells and manual determination of precise cell positions.ResultsWe have quantitatively analyzed these error sources, demonstrating that manual cell tracking of pancreatic cancer cells lead to mis-calculation of migration rates of up to 410%. In order to provide for objective measurements of cell migration rates, we have employed multi-target tracking technologies commonly used in radar applications to develop fully automated cell identification and tracking system suitable for high throughput screening of video sequences of unstained living cells.ConclusionWe demonstrate that our automatic multi target tracking system identifies cell objects, follows individual cells and computes migration rates with high precision, clearly outperforming manual procedures.
Highlights
Cell motility is a critical parameter in many physiological as well as pathophysiological processes
Using migration of different populations of pancreatic cancer cells as a model system, we show that the results of manual cell tracking are highly variable and lead to mis-calculation of migration rates by up to 410%
Variability of cell speed estimation caused by manual centroid selection The migration rate of cells is commonly measured via the mean displacement (MD, i.e. the mean distance traveled per minute) of the cell centroid
Summary
Cell motility is a critical parameter in many physiological as well as pathophysiological processes. Cell motility is critical in processes such as wound healing, which requires fibroblasts and keratinocytes to migrate into wound sites [3,4], or the immune response, which involves extensive migratory activity of immune cells to and from lymphoid tissues and distant sites of infection [5,6,7,8]. In addition to these physiological roles, cell migration is an important parameter in pathological processes such as carcinogenesis. Using migration of different populations of pancreatic cancer cells as a model system, we show that the results of manual cell tracking are highly variable and lead to mis-calculation of migration rates by up to 410%
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