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March 01, 2009 (Vol. 29, No. 5)

Measuring Multiplexed Signal Transduction

Amnis’ Platform Was Developed to Assist in Assessment of NF-kappa-B Translocation

  • The common goal of many drug discovery efforts is the modulation of immune responses, for example by the inhibition of tumor necrosis factor-alpha (TNF-a). TNF-a is a potent activator of many cells of the immune system and plays a large role in the pathogenesis of asthma and of autoimmune diseases such as rheumatoid arthritis. TNF-a mediates its pro-inflammatory effect by inducing translocation of NF-kB from the cytoplasm to the nucleus.

    In this article, we present a multiplexed method for the measurement of NF-kB translocation in THP1 cells, a monocytic cell line, in response to TNF-a stimulation at various doses and exposure times. The Amnis ImageStream platform employed in this study images cells directly in flow, making it well-suited to the analysis of both primary and cultured immune cells. Imaging flow cytometry generates over 100,000 cell images per minute to achieve high assay robustness even with rare sub-populations.

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