Cell Motility Reagent Kit

• Versatile assay is widely applicable to a variety of cell types
• Validated protocol for sample preparation/processing requires 2.5 hours post-compound incubation
• Assay utilizes fluorescence reagents and requires no cell lysis, purification or filtration steps; and no handling of radioactive isotopes
• Cells prepared with the HCS Reagent Kits may be analyzed directly via standard fluorescence microscopy; or any HCS imaging systems

The Cell Motility HCS Reagent Kit provides reagents and a protocol optimized for quantification of cell motility by directly measuring the size of tracks generated by migrating cells. The assay is performed on live cells growing on standard high density microplates. The kit includes Blue Fluorescent Beads, Rhodamine-conjugated Phalloidin for staining the cytoskeleton and assay buffers.

Cell motility is central to a number of biological and pathologic processes including cancer cell invasion and metastasis, inflammation, angiogenesis, wound repair, and embryonic development. The movement of cells occurs via the concerted activities of cell adhesion molecules, the actin cytoskeleton and an extensive network of signaling molecules. A combination of cell substrate adhesion, cell spreading, lamellar protrusion and directional control work together for net translocation. These activities are regulated both by external factors including extracellular matrix proteins, cytokines and soluble growth factors, and by intracellular signal transduction cascades.

The Cell Motility Kit facilitates the quantification of cell movement with images of fixed cells. This functional assay can predict the efficacy of potential drugs for a number of therapeutic areas. The assay is performed by plating cells on a lawn of microscopic fluorescent beads. As cells move across the lawn, they phagocytose and push aside the beads, clearing phagokinetic tracks behind them. The track area is proportional to the magnitude of cell movement. The reagents in this kit combined with the ArrayScan HCS Reader and the Cell Motility BioApplication Software enable automated plate handling, focusing, cell image acquisition, analysis and quantification. For a more detailed description of the image-processing algorithm, see the Cell Motility BioApplication Guide that accompanies the Cell Motility BioApplication software.

Figure 1. Images of tracks (black) generated by A549 cells in the absence (unstimulated, left) or presence (stimulated, right) of serum. Cells appear as red fluorescent spots due to the cytoplasmic staining (rhodamine-conjugated phalloidin). Images were acquired by the ArrayScan HCS Reader.

Figure 2. Dose response curve for cell motility stimulation in L9292 cells.

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