ProtocolImaging

Simultaneous Optical Measurements of Cytosolic Ca2+ and cAMP in Single Cells

Science's STKE  19 Sep 2006:
Vol. 2006, Issue 353, pp. pl6
DOI: 10.1126/stke.3532006pl6

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Abstract

Understanding the temporal and spatial integration of the Ca2+ and adenosine 3′,5′-monophosphate (cAMP) signaling pathways requires concurrent measurements of both second messengers. Here, we describe an optical technique to simultaneously image cAMP and Ca2+ concentration gradients in MIN6 mouse insulinoma cells using Epac1-camps, a Förster (or fluorescence) resonance energy transfer (FRET)-based cAMP biosensor, and Fura-2, a fluorescent indicator of Ca2+. This real-time imaging method allows investigation of the dynamic organization and integration of multiple levels of signal processing in single living cells.

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