The authors have developed a cellular analysis platform, based on encoded microcarriers, that enables the multiplexed analysis of a diverse range of cellular assays. At the core of this technology are classes of microcarriers that have unique, identifiable codes that are deciphered using CCD-based imaging and subsequent image analysis. The platform is compatible with a wide variety of cellular imaging-based assays, including calcium flux, reporter gene activation, cytotoxicity, and proliferation. In addition, the platform is compatible with both colorimetric and fluorescent readouts. Notably, this technology has the unique ability to multiplex different cell lines in a single microplate well, enabling scientists to perform assays and data analysis in novel

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Image cytometry has made possible the collection and analysis of multiparameter cellular information. The wider use of image cytometry in drug screening will depend on its throughput, efficiency, repeatability, and on the added benefits compared with less sophisticated but faster methods. Throughput (number of datapoints per unit of time) and efficiency (number of datapoints from the given amount of reagents or plate area) are addressed here by screening multiple cell lines simultaneously using encoded carriers (CellCards®). CellCards are rectangular particles with an expandable color barcode and a transparent section for cellular readout. Before performing the assay, each cell line is grown on a different class of carriers. CellCards, with attached cells, are mixed and dispensed into a microtiter plate where the assay is performed. Next the plates are imaged, decoded and the cells associated with each CellCard class are analyzed. Using CellCards the efficiency is increased by the multiplexing factor (the number of cell lines analyzed in each well). We routinely run assays with a multiplex factor of ten. Throughput is additionally addressed by working at the lowest possible magnification for a given assay. Decoding of CellCards requires one image per well in 96-well microtiter plate format. The system provides the added benefit of internal consistency since the data can be normalized to controls within each well.

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The desire to obtain more biologically relevant data is expanding the use of cell-based assays in drug discovery. These assays are performed and analyzed in ever more sophisticated ways (e.g. high content screening) that allow the collection of multiparametric information about cells affected by the screened compounds. The driver for these developments is the desire to increase data quality and density and reduce the use of valuable reagents and time.Here we describe an approach that adds a new dimension to the data quality/quantity mix by simultaneously analyzing several cell types in the same microplate well. The system is based on the use of encoded carriers (CellCards®) that permit the reading and analysis of cellular responses, and at the same time allow decoding and the attribution of these responses to the appropriate cell line. CellCards are rectangular particles with an expandable color barcode and a transparent section upon which cells can be grown and imaged for cellular readout. Before performing the assay, each cell line is grown on a different class of CellCards. CellCards, with attached cells, are mixed and dispensed into a microtiter plate where the assay is performed. Next the plates are imaged, decoded and the cells associated with each CellCard class are analyzed.Multiplexing cell lines allows assay controls and data normalization within each well, reducing well-to-well variability. It also allows the simultaneous interrogation of multiple targets and thus concurrent potency and selectivity screening. This may significantly reduce the time required to take a compound from primary screening into the clinic.