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Exposing Cells to a Single Pulse of Light of user-defined Wavelength and Duration using the FDSS6000

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Exposing Cells to a Single Pulse of Light of user-defined Wavelength and Duration using the FDSS6000

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In the described model system the GPCR receptor optimally transduces signal from light at 480 nm (light as a wave) by absorbing photon energy (light as a particle) and undergoing isomerization. As a consequence the excitation wavelength 480 nm used for Fluo-4 AM based calcium dyes also agonizes the receptor. In this model the assay design. Precludes collecting a pre-agonist (background) signal and Continuously bombards the receptor with stimulus. In this study we used. The UV ratiometric dye Fura-2 AM to separate reporter dye excitation/emission from receptor agonism and. A novel "pulse" feature in the FDSS6000 software to expose cells for specific times to 480 nm light. The result is an assay that can collect background signal and determine the minimal light exposure time at 480 nm (or any other wavelength) for maximal agonism.

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