( ay-specific application modules design...)High-content screening of GPCR activation with MetaXpress, AcuityXpress and the Transfluor assay system

ay-specific application modules, designed for easy, interactive optimization of segmentation parameters. The simplified interface allows simple configuration and testing of analysis settings for your particular assay.(See Figure 3, Panel A.)

The MetaXpress Transfluor Module uses the proprietary Adaptive Background Correction system to distinguish pits, vesicles and nuclei from their local surroundings so they can be selectively measured and counted, even in the presence of uneven staining or protein expression levels. This can be difficult to do using ordinary thresholding methods because the intensity of the pits can be marginally above background, and the GFP-tagged beta-arrestin expression levels can vary from cell to cell.

With the Transfluor Module, designed for cell-by-cell analysis of Transfluor data, pits and vesicles can be detected and distinguished simultaneously. The optional nuclear detection provides accurate counting of cells for data normalization. Three user-adjustable parameters are utilized for the image analysis. The Approximate min width and Approximate max width parameters are modified as needed for the expected size of the pits, vesicles or nuclei. The Intensity above local background parameter is adjusted for optimal discrimination of each pit or vesicle from the local background. This significantly improves the ability to identify and accurately quantitate actual pit and vesicle formation even if the intensity varies between cells in the image.

Multiple measurements can be logged to the database, including the number of pits or vesicles, total area covered by pits or vesicles, average and integrated intensity of the granules, and nuclear area and intensity. Results
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