Characterization of single-cell migration using a computer-aided fluorescence time-lapse videomicroscopy system.

Abstract

Single-cell assays of cell migration, while yielding dynamic measurements of cell position and morphology, are predominantly limited by the time required for data collection and analysis. Computer-aided fluorescence time-lapse videomicroscopy (CAFTiV) was developed in order to facilitate the tracking and rapid examination of large numbers of motile cells. The system combines time-lapse videomicroscopy with epifluorescence capability, which allows full automation of image capture, sorting, and analysis due to the low background in the fluorescence images. Utilizing the CAFTiV system, data analysis time was reduced from over 125 h to less than 1 labor minute. In addition, fluorescence imaging permits cell tracking in small-volume chambers (<100 microL), which is useful should the addition of expensive reagents be required. It is anticipated that the ability to characterize both biochemical and biophysical properties responsible for cell movement will be enhanced by this methodology.