engleski

A system for monitoring and analyzing cell motility

Cell motility plays a crucial role in numerous physiological and pathological processes, including embryonic development, immune response, neurite outgrowth, wound healing, and metastasis of tumor cells. Chemotaxis of cells is governed by elaborate biomolecular cascades that cells utilize to detect an external signal and move toward it. In order to understand basic principles of cell motility, migration of cells is often studied in the absence of external signals. By investigating patterns of movement apparent on macroscopic scale, an insight into underlying molecular mechanisms that govern the cell migration can be gained. For instance, mechanisms controlling extension of the pseudopod, traction between a cell and the substratum, and retraction of the rear cell end can be studied in this way. We have assembled a system suitable for monitoring and analyzing migration of a few dozen small-sized eukaryotic cells. Movement of cells is monitored either by phase-contrast microscopy or by dark-field microscopy, depending on the requirements of the tracking experiment. Images acquired at a constant time interval are stored and subsequently analyzed with an appropriate image processing software package. Result of the image processing are cell tracks that are presented as time sequences of cell centroid coordinates. These numerical data are further analyzed in order to get information on parameters that quantitatively describe cell motility such as speed and directionality. Directionality of movement may have a major impact on how cells colonize a new environment, e.g. during food seeking, morphogenesis and metastasis. We are currently using this system to investigate spatio-temporal patterns that occur during the movement of Dictyostelium discoideum cells in an isotropic environment. The system can also be used in analyzing cell motility of other motile eukaryotic cells of similar size, e.g. neutrophils. To monitor long-term movement of human cell lines, it would be necessary to provide cells with optimal conditions, for example by placing them into a chamber with the controlled temperature and atmosphere.

cell motility

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