Abstract
In multi-cellular organisms, cell-cell communication involves a large variety of cell-surface receptors that bind specific ligands. Unraveling these cell signaling networks in depth requires quantitative information on ligand-receptor interactions within living systems. Fluorescence correlation spectroscopy (FCS) based methods are powerful approaches to obtain these data. Still, their application to living systems have proven challenging. We have developed an integrated dual-color dual-focus line-scanning fluorescence correlation spectroscopy (2c2f lsFCS) technique that greatly facilitates live-cell and tissue experiments. Ligand and receptor concentrations as well as their diffusion coefficients within the cell membrane can be quantified without performing additional calibration experiments. In a first application, we have employed this robust technique to study interactions between Wnt antagonists, Dickkopf1 and Dickkopf2 (Dkk1/2), and their cognate receptor, low-density-lipoprotein-receptor related protein 6 (LRP6), in the plasma membrane of HEK293T cells.
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