Abstract
Understanding how cells migrate individually and collectively during development and cancer metastasis can be significantly aided by a computation tool to accurately measure not only cellular migration speed, but also migration direction and changes in migration direction in a temporal and spatial manner. We have developed such a tool for cell migration researchers, named Pathfinder, which is capable of simultaneously measuring the migration speed, migration direction, and changes in migration directions of thousands of cells both instantaneously and over long periods of time from fluorescence microscopy data. Additionally, we demonstrate how the Pathfinder software can be used to quantify collective cell migration. The novel capability of the Pathfinder software to measure the changes in migration direction of large populations of cells in a spatiotemporal manner will aid cellular migration research by providing a robust method for determining the mechanisms of cellular guidance during individual and collective cell migration.
Highlights
Cellular migration has been shown to be an important process in cancer progression, development, tissue repair, and immune response [1–10]
We have developed an automated high throughput cell tracking software, named Pathfinder, which is capable of simultaneously measuring and reporting cellular migration speed, migration direction and changes in migration direction of thousands of fluorescently labeled cells for an unlimited number of microscopy videos
Transforming Growth Factor-Beta (TGFb) stimulations were conducted with 100 pM ligand, while Epidermal Growth Factor (EGF) stimulations were conducted with 100 nM ligand
Summary
Cellular migration has been shown to be an important process in cancer progression, development, tissue repair, and immune response [1–10]. A plethora of research has been performed to identify the molecular mechanisms behind how individual cells achieve migration, as well as how neighboring cells migrate cooperatively in collective migration (reviewed in [11–13] and [14], respectively). The collective migration of cells has been shown to be an important process in cancer progression, development and wound repair [16–23]. Such collective behavior results from each cell responding to the environmental stimuli of neighboring cells, in addition to non-cell environmental stimuli [4,5,14,15,17,19,20,24–32]. A relatively large amount of research has been conducted to determine mechanisms behind individual cell migration, far less is known about exactly how cells migrate collectively. There is no standard method in the literature to quantify the ‘collectiveness’ behavior during collective migration [33–35]
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