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. 2006 Aug 8;51(1):7–19. doi: 10.1007/s10616-006-9006-7

Automated time-lapse microscopy and high-resolution tracking of cell migration

Joseph S Fotos 1, Vivek P Patel 1, Norman J Karin 1,3, Murali K Temburni 1,2, John T Koh 2, Deni S Galileo 1,✉
PMCID: PMC3449480  PMID: 19002890

Abstract

We describe a novel fully automated high-throughput time-lapse microscopy system and evaluate its performance for precisely tracking the motility of several glioma and osteoblastic cell lines. Use of this system revealed cell motility behavior not discernable with conventional techniques by collecting data (1) from closely spaced time points (minutes), (2) over long periods (hours to days), (3) from multiple areas of interest, (4) in parallel under several different experimental conditions. Quantitation of true individual and average cell velocity and path length was obtained with high spatial and temporal resolution in “scratch” or “wound healing” assays. This revealed unique motility dynamics of drug-treated and adhesion molecule-transfected cells and, thus, this is a considerable improvement over current methods of measurement and analysis. Several fluorescent vital labeling methods commonly used for end-point analyses (GFP expression, DiO lipophilic dye, and Qtracker nanocrystals) were found to be useful for time-lapse studies under specific conditions that are described. To illustrate one application, fluorescently labeled tumor cells were seeded onto cell monolayers expressing ectopic adhesion molecules, and this resulted in consistently reduced tumor cell migration velocities. These highly quantitative time-lapse analysis methods will promote the creation of new cell motility assays and increase the resolution and accuracy of existing assays.

Keywords: Cell migration, Green fluorescent protein, Scratch assay, Time-lapse, Tumor cell lines, Vital fluorescent labeling

Full Text

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Acknowledgements

This work was supported by grants from NIH to D.S.G. (NS040317), to J.K. (NS049523), and to N.J.K. (HD042066). We gratefully acknowledge that the filter cube with custom excitation bandpass filters was provided by Chroma Technology Corporation. J. Fotos and V. Patel were awarded undergraduate research fellowships through grants to the Univ. of Delaware from the Howard Hughes Medical Institute and the Ronald E.␣McNair Program. We gratefully thank Dr. John Kappes for the GFP lentiviral vector construct and Dr. Marty Grumet for the NgCAM/L1 cDNA construct.

Footnotes

Joseph S. Fotos and Vivek P. Patel contributed equally to this work

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