Automated time-lapse microscopy and high-resolution tracking of cell migration
The study of cell motility is greatly enhanced by using a fully-automated high-throughput time-lapse microscopy system that is capable of collecting and analyzing data (1) from closely-spaced time points (seconds to minutes), (2) over long periods (hours to days), (3) from multiple areas of interest, (4) under several different experimental conditions simultaneously. Time-lapse video images collected under phase contrast and fluorescent illumination were analyzed using parameters of migration velocity, total accumulated distance (path length), and directionality for individual cells or for averaged cell populations. Quantitation of ''scratch'' or ''wound healing'' assays revealed unique motility dynamics of drug-treated and adhesion molecule-transfected cells with high resolution and, thus, is a vast distinct improvement of current methods. Several fluorescent vital labeling methods commonly used for end-point analyses, including GFP expression, were evaluated and most were useful for time-lapse studies under specific conditions. For example, fluorescently-labeled tumor cells were seeded onto cell monolayers expressing ectopic adhesion molecules displayed altered migration velocities compared to tumor cells plated directly onto culture dishes. The techniques described here revealed cell motility behavior not discernable by previously-used methods. We propose that quantitative time-lapse video analysis will foster the creation new cell motility assays, and increase the resolution and accuracy of existing assays.
- Research Organization:
- Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 891413
- Report Number(s):
- PNNL-SA-46775; TRN: US200621%%780
- Journal Information:
- Cytotechnology, 51(1):7-19, Journal Name: Cytotechnology, 51(1):7-19
- Country of Publication:
- United States
- Language:
- English
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