Related Experiment Video
Updated: Nov 23, 2025
Simultaneous Imaging of Microglial Dynamics and Neuronal Activity in Awake Mice
Published on: August 23, 2022
A Protocol for FRET-Based Live-Cell Imaging in Microglia
Renato Socodato1, Pedro Melo1, José P Ferraz-Nogueira1
1Instituto de Investigação e Inovação em Saúde and Instituto de Biologia Molecular e Celular, Universidade do Porto, 4200-135 Porto, Portugal.
Abstract:
This protocol highlights the use of FRET-based biosensors to investigate signaling events during microglia activation in real time. Understanding microglia activation has gained momentum as it can help decipher signaling mechanisms underlying the neurodegenerative process occurring in neurological disorders. Unlike more traditional methods widely employed in the microglia field, FRET allows microglia signaling events to be studied in real time with exquisite subcellular resolution. However, FRET-based live-cell imaging requires application-specific biosensors and specialized imaging systems, limiting its use in in vivo studies. For complete details on the use and execution of this protocol, please refer to Socodato et al. (2020), Portugal et al. (2017), and Socodato et al. (2018).
Insights
This protocol uses Förster Resonance Energy Transfer (FRET) biosensors to study microglia activation in real time. This method offers high subcellular resolution for understanding neurological disorders.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Microglia activation is crucial for understanding neurological disorders.
- Traditional methods lack real-time, high-resolution insights into microglia signaling.
- Förster Resonance Energy Transfer (FRET) offers a novel approach.
Purpose of the Study:
- To present a protocol for using FRET-based biosensors.
- To investigate real-time signaling events during microglia activation.
- To enhance understanding of neurodegenerative processes.
Main Methods:
- Utilizing FRET-based biosensors for live-cell imaging.
- Applying specialized imaging systems for microglia studies.
- Real-time monitoring of subcellular signaling events.
Main Results:
- Demonstrated the capability of FRET to capture dynamic microglia signaling.
- Achieved exquisite subcellular resolution in observing activation events.
- Provided a protocol for detailed mechanistic insights.
Conclusions:
- FRET-based biosensors are powerful tools for studying microglia activation.
- This technique advances the understanding of signaling in neurological disorders.
- Limitations include the need for specific biosensors and imaging systems for in vivo studies.
