Automated Imaging-Based Technique for Monitoring of Ca2+ and DAG Biosensors Enables Detailed Characterization of GPCR Kinetics

This poster describes an automated imaging-based approach to characterize the IP₃ /DAG signaling pathway using multiplexed R-GECO Ca²⁺ and DAG fluorescent biosensors from Montana Molecular. This method produces detailed kinetic profiles of Ca²⁺ flux and DAG levels following activation of G_q-coupled receptors within the same population of cells. Background subtraction and image analysis tools enable detection of changes in Ca²⁺ and DAG levels that are more sensitive than techniques relying on bulk fluorescence measurements. Additionally, this method enables single-cell analysis for detailed characterization of response dynamics and subcellular effects.