These stains are intense, highly photostable and match the output wavelengths of common fluorescence instrumentation. Effective image analysis in HCS cell-based assays and fluorescence microscopy requires fluorescent labeling of the entire cell. In these assays, the cellular primary object is used to identify and count individual cells and define the cell region in which the image analysis is applied. The primary object might be a major cellular component, such as the nucleus, a large organelle or the whole cell. When the whole cell is the primary object, high-quality Whole Cell Stains effectively distinguish intact cells from bordering cells when analyzed by appropriate software.
Figure 1. A549 cells stained with BrdU antibody and DyLight 488 secondary antibody (left) and Whole Cell Stain Red for 30 min (center). Overlay (right). Cell images were acquired on an ArrayScan HCS reader with a 20X objective lens. Whole Cell Stain Red does not interfere with BrdU detection.
Figure 2. Sub-confluent NIH 3T3 cells stained with Whole Cell Stain Blue and 7-AAD (nuclear stain) for 15 min. Cell images were acquired on an ArrayScan HCS reader with a 20X objective lens, and individual cells in the image were identified using the Cellomics Morphology Explorer BioApplication; cell-to-cell separation was achieved using the Watershed object segmentation algorithm option in the Morphology Explorer BioApplication. Similar Cell Separation Index measurements made with Whole Cell Stain Green, Orange, and Red using DAPI as a nuclear stain also provide good cell-to-cell separation.
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