Fret Calcium Sensor
Fret Calcium Sensor - Related Videos
09:12
G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay
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Cited by 4 •
2016
Simple methods to detect the selective activation of G proteins by G protein-coupled receptors remain an outstanding challenge in cell signaling. Here, Fӧrster resonance energy transfer (FRET) biosensors have been developed by pairwise tethering a GPCR to G protein peptides to probe conformational changes at controlled concentrations in live cells.
09:21
Real Time Monitoring of Intracellular Bile Acid Dynamics Using a Genetically Encoded FRET-based Bile Acid Sensor
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Cited by 8 •
2016
We provide a detailed protocol to study bile acid dynamics in living cells using a genetically encoded BAS FRET sensor. This Bile Acid Sensor represents a unique tool to study (regulation of) bile acid transport and FXR activation in a wide range of cell types.
03:39
In Vivo Calcium Imaging Using Synaptically Localized Calcium Sensors in Drosophila melanogaster
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2025
This video demonstrates a method for in vivo calcium imaging using synaptically localized calcium sensors in flies. It outlines the steps for preparing the flies for microscopy, conducting olfactory conditioning, and visualizing odor-induced calcium influx.
03:40
Imaging Intracellular ATP Dynamics Using FRET-Based Sensors in an Organotypic Mouse Hippocampal Slice
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2025
Source: Lerchundi, R., et. al. Imaging of Intracellular ATP in Organotypic Tissue Slices of the Mouse Brain using the FRET-based Sensor ATeam1.03YEMK. J. Vis. Exp. (2019)This video demonstrates the use of FRET(Fluorescence Resonance Energy Transfer)-based sensors to measure intracellular ATP levels in organotypic mouse hippocampal slices, enabling real-time visualization of ATP dynamics in neurons and astrocytes under physiological and experimental conditions.
12:30
Monitoring ER/SR Calcium Release with the Targeted Ca2+ Sensor CatchER+
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Cited by 11 •
2017
We present protocols for the application of our targeted genetically-encoded calcium indicator (GECI) CatchER+ for monitoring rapid calcium transients in the endoplasmic/sarcoplasmic reticulum (ER/SR) of HEK293 and skeletal muscle C2C12 cells using real-time fluorescence microscopy. A protocol for the in situ Kd measurement and calibration is also discussed.
FAQs
Calcium binding changes the conformation of the sensing domain, which modifies the relationship between the fluorescent donor and acceptor. That structural change alters Förster resonance energy transfer and, consequently, the measured fluorescence ratio. The probe therefore converts a calcium-dependent molecular event into an optical signal that can be tracked as neuronal activity changes over time.
Comparing fluorescence as a ratio provides a quantitative basis for comparing calcium signals across experiments, rather than relying only on one fluorescence measurement. This is especially useful when investigators examine dynamic neural processes, because the ratio reflects changes in the sensor’s energy-transfer state while supporting comparisons among measurements of neuronal activity.
The sensing domain links calcium binding to the optical behavior of the probe. When its conformation changes, the relative interaction between the fluorescent donor and acceptor changes as well, modifying energy transfer and the fluorescence ratio. This coupling is what allows intracellular calcium variation to be represented as a measurable change in fluorescence.
Investigators image neurons or neural circuits with the sensor and monitor changes in the fluorescence ratio over time. They can then examine how calcium dynamics correspond to neuronal activity, synaptic input, electrical activity, or cellular signaling. This workflow turns a time-varying optical measurement into a way to study neural processes as they occur.
Real-time measurements allow researchers to follow calcium dynamics while relating them to synaptic input, electrical activity, and downstream cellular signaling. The fluorescence ratio supplies a quantitative signal for examining these relationships in neurons and neural circuits. This makes the approach useful for studying how activity-associated calcium changes are expressed within neural systems.
In neuroscience, these sensors can be used to investigate neural activity at the level of individual neurons and neural circuits, and to connect cellular signals with behavior or disease. Their ability to report changing calcium levels in real time supports studies of dynamic neural processes rather than only static cellular states.