Cell Viability Reagent Kit

• Assay is versitle and widely applicable to a variety of cell types
• Utilizes fluorescent 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 HCS imaging systems.
• DeadDye™ and VitalDye™ fluorescent probes offer sensitivite, quantitative and qualitative screening results without radioactivity   

The Cellomics Cell Viability Reagent Kit for high-content screening enables quantification of live and dead cells, and total cell counts. The kit provides a fixed end-point assay based on fluorescence detection of live and dead cells distributed on microplates. The core reagents supplied include DeadDye and VitalDye. The nuclei are identified by DNA-specific Hoechst Dye, which is also included in the kit.

When cells are dead or dying, plasma membranes leak and become permeable to large macromolecules. In this state, DeadDye enters the cell and is visualized by its red fluorescence emission. VitalDye, a membrane-permeable fluorescent probe, operates as a live-cell indicator. Soluble cytoplasmic enzymes cleave VitalDye, trapping its fluorescent moiety inside intact cells. Live cells stained with VitalDye are distinguished from dead cells by their green fluorescence emission. Identification of the entire cell population is achieved using Hoechst Dye, a membrane-permeable nucleic acid stain emitting in the blue fluorescence channel. DeadDye and VitalDye may be used separately or in combination to quantify dead cells, live cells or both cell populations.

The Cell Viability Kit Reagents, in combination with the ArrayScan HCS System and the Cell Viability Application software enable automated plate handling, focusing, cell image acquisition, and analysis and cell viability quantification. For a more detailed description of the image processing algorithm, see the Cell Viability Application Guide that accompanies the software.

Figure 1. Live and dead cells distinguished by the ArrayScan HCS Reader and the Cell Viability Kit Reagents. HeLa cells were plated in a 96-well microplate and then treated with the Positve Control Compound (supplied with the kit). Hoechst staining identifies all cells in the imaged field (A). VitalDye (green) and DeadDye (red) show live and dead cells, respectively (B).

Figure 2. Dose response curve of HeLa cells treated with the Positive Control Compound.

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