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Measurement of 3-Dimensional cAMP Distributions in Living Cells using 4-Dimensional (x, y, z, and λ) Hyperspectral FRET Imaging and Analysis
 

Measurement of 3-Dimensional cAMP Distributions in Living Cells using 4-Dimensional (x, y, z, and λ) Hyperspectral FRET Imaging and Analysis

Article DOI: 10.3791/61720-v • 08:22 min • October 27th, 2020
October 27th, 2020 •
1,2, 3, 1, 3, 1,2, 1,2, 1,2,3
1Department of Pharmacology, University of South Alabama, 2Center for Lung Biology, University of South Alabama, 3Department of Chemical and Biomolecular Engineering, University of South Alabama

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Due to inherent low signal-to-noise ratio (SNR) of Fӧrster resonance energy transfer (FRET) based sensors, measurement of cAMP signals has been challenging, especially in three spatial dimensions. Here, we describe a hyperspectral FRET imaging and analysis methodology that allows measurement of cAMP distribution in three spatial dimensions.

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Keywords: 3D CAMP Distribution 4D Hyperspectral FRET Imaging Cyclic AMP FRET Confocal Microscopy Spectral Imaging 3D Cell Imaging Live Cell Imaging Tyrodes Buffer Nuclear Labeling Laser Settings Z-stack Acquisition
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