Open AccessIndividual cell motility studied by time‐lapse video recording: Influence of experimental conditions
Author(s) -
HartmannPetersen Rasmus,
Walmod Peter S.,
Berezin Anton,
Berezin Vladimir,
Bock Elisabeth
Publication year - 2000
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/1097-0320(20000801)40:4<260::aid-cyto2>3.0.co;2-j
Subject(s) - motility , cell , microbiology and biotechnology , biophysics , video microscopy , actin , cell culture , confocal microscopy , confocal laser scanning microscopy , cell growth , biology , seeding , morphology (biology) , chemistry , biochemistry , agronomy , genetics
Background Eukaryotic cell motility plays a key role during development, wound healing, and tumour invasion. Computer‐assisted image analysis now makes it a realistic task to quantify individual cell motility of a large number of cells. However, the influence of culture conditions before and during measurements has not been investigated systematically. Methods We have evaluated intraassay and interassay variations in determinations of cellular speed of fibroblastoid L929 cells and investigated the effects of a series of physical and biological parameters on the motile behavior of this cell line. Cellular morphology and organization of filamentous actin were assessed by means of phase‐contrast and confocal laser scanning microscopy and compared to the corresponding motility data. Results Cell dissociation procedure, seeding density, time of cultivation, and substrate concentration were shown to affect cellular speed significantly. pH and temperature of the medium most profoundly influenced cell motility and morphology. Thus, the mean cell speed was 40% lower at pH 7.25 than at pH 7.6; at 29°C, it was approximately four times lower than at 39°C. Conclusion Of the parameters evaluated, cell motility was most strongly affected by changes in pH and temperature. In general, changes in cell speed were accompanied by alterations in cell morphology and organization of filamentous actin, although no consistent phenotypic characteristics could be demonstrated for cells exhibiting high cell speed. Cytometry 40:260–270, 2000 © 2000 Wiley‐Liss, Inc.