We have developed a solution for cell motility measurements with the Cell Motility BioApplication and HCS Reagent Kit. These tools work together to provide an automated method for quantifying cell motility in response to drug candidates. The kit provides High Content Screening (HCS) quality fluorescence reagents and a protocol optimized for visualizing and quantifying cell movement by directly measuring the size of tracks generated by migrating cells. It provides a functional assay that can predict the efficacy of potential drugs for a number of therapeutic areas. The assay is performed on live cells plated on a lawn of microscopic fluorescent beads. As cells move across the lawn, they phagocytose the beads, clearing phagokinetic tracks behind them (Figure 1). The track area is proportional to the magnitude of cell movement. Included in the kit are buffers needed for processing the assay, blue fluorescent microspheres, and rhodamine-conjugated phalloidin.

The BioApplication identifies the tracks produced by motile cells based on intensity of the fluorescent lawn of beads and measures the area of the tracks produced by these cells (Figure 2). It also identifies cells associated with the tracks based on the cytoskeletal stain of the cells and quantifies the area of the cells. The BioApplication does not process tracks that do not contain cells. Cells or small colonies are identified as a single object if their cytoplasm is contiguous. Thus, cells are characterized by the total area of their lamellipodia. The user can specify an upper limit on the number of cells per track, thereby enabling analysis of single cell tracks if desired.

(Figure 3) below shows the cell motility response of L929 fibroblast cells to increasing concentrations of serum. This data demonstrates that cell motility can be measured in a quantitative, objective, and streamlined manner, resulting in improved workflow for studying the effects of drug candidates on cell motility.