Journal
CYTOTECHNOLOGY
Volume 51, Issue 1, Pages 7-19Publisher
SPRINGER
DOI: 10.1007/s10616-006-9006-7
Keywords
cell migration; green fluorescent protein; scratch assay; time-lapse; tumor cell lines; vital fluorescent labeling
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Funding
- NICHD NIH HHS [R03 HD042066] Funding Source: Medline
- NINDS NIH HHS [R21 NS040317, R01 NS049523] Funding Source: Medline
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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.
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