Presentation: P03049

Although most luminescent reporters methods are destructive to cell samples, intracellular luminescence from living cells can be measured by adding substrates to the growth medium. We have previously reported the ability to simultaneously measure two luminescent signals. We now report an extension of the color-separation between the luciferases in living cells by using the blue emission of Renilla luciferase (480 nm) with the red emission of the Chroma-Luc™ luciferase (611 nm) The combined light emission can be achieved by adding a mixture of EnduRen™ Live Cell Substate and beetle luciferin to the culture medium. We have demonstrated the advantages of multiplexed reporter measurements using isoproterenol and inhibitors with CRE-element, TNF-α with NFkB, doxycycline with Tet-off system, toxins with housekeeping promoters. For example, Z’ increased from 0.24 to 0.70 when Renilla luminescence controlled by the CRE-element in living cells is normalized to red Chroma-luc™ luminescence controlled by the SV40 promoter, measured concurrently.

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