1 July 2004 Multiplexed high-throughput image cytometry using encoded carriers
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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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Ilya Ravkin, Vladimir Temov, Aaron D. Nelson, Michael A. Zarowitz, Matthew Hoopes, Yuli Verhovsky, Gregory Ascue, Simon Goldbard, Oren Beske, Bhagyashree Bhagwat, Holly Marciniak, "Multiplexed high-throughput image cytometry using encoded carriers", Proc. SPIE 5322, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues II, (1 July 2004); doi: 10.1117/12.528071; https://doi.org/10.1117/12.528071


Image analysis

Image segmentation



Quality measurement

Control systems

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