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Updated: Jun 5, 2026

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

Published on: August 20, 2012

Dynamic visualization of physiological CaMKII activity using sensitive FRET biosensors.

Sohum Mehta¹, Nidhi A Thaker², Kengo Adachi³

¹Department of Pharmacology, University of California San Diego, La Jolla, CA 92093, USA.

Biorxiv : the Preprint Server for Biology

|June 4, 2026

Summary

This summary is machine-generated.

Researchers developed new biosensors to visualize Calcium-calmodulin (CaM)-dependent protein kinase II (CaMKII) activity. These tools enable sensitive and specific monitoring of CaMKII signaling in living cells and tissues, advancing health and disease research.

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Last Updated: Jun 5, 2026

FRET Microscopy for Real-time Monitoring of Signaling Events in Live Cells Using Unimolecular Biosensors

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Published on: August 20, 2012

Preclinical Cardiac Electrophysiology Assessment by Dual Voltage and Calcium Optical Mapping of Human Organotypic Cardiac Slices

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Designing a Bioreactor to Improve Data Acquisition and Model Throughput of Engineered Cardiac Tissues

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Published on: June 2, 2023

Area of Science:

Molecular Biology
Neuroscience
Biochemistry

Background:

Calcium-calmodulin (CaM)-dependent protein kinase II (CaMKII) regulates vital physiological processes.
Visualizing CaMKII activity in living systems is crucial for understanding health and disease.
Current methods lack the sensitivity and specificity for robust CaMKII monitoring.

Purpose of the Study:

To engineer novel Förster resonance energy transfer (FRET)-based biosensors for CaMKII.
To achieve high specificity, sensitivity, and signal-to-noise ratio for CaMKII activity reporters.
To enable quantitative visualization of endogenous CaMKII signaling in various cell types and tissues.

Main Methods:

Leveraged a serine/threonine kinome-wide substrate atlas for rational biosensor design.
Engineered a suite of FRET-based CaMKII kinase activity reporters.
Utilized 2-photon fluorescence lifetime imaging (2pFLIM) on organotypic hippocampal slices.

Main Results:

Developed highly specific and sensitive FRET biosensors for CaMKII.
Successfully visualized endogenous CaMKII activity in cultured cell lines, cardiomyocytes, oocytes, and neurons.
Quantitatively tracked LTP-induced CaMKII activity in single dendritic spines using 2pFLIM imaging.

Conclusions:

The new FRET biosensors represent a significant advance for studying CaMKII signaling.
These tools allow for robust and quantitative monitoring of CaMKII dynamics in physiologically relevant contexts.
Enables deeper insights into the molecular regulation of CaMKII in health and disease.