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Fast high-resolution miniature two-photon microscopy for brain imaging in freely behaving mice
Weijian Zong1,2, Runlong Wu1, Mingli Li1
1State Key Laboratory of Membrane Biology, Institute of Molecular Medicine, Peking-Tsinghua Center for Life Sciences, Beijing Key Laboratory of Cardiometabolic Molecular Medicine, Peking University, Beijing, China.
Nature Methods
|May 30, 2017
Summary
Researchers developed a novel miniaturized two-photon microscope for high-resolution imaging of neuronal activity at single dendritic spines in freely moving animals. This breakthrough allows detailed study of brain function during natural behaviors.
Area of Science:
- Neuroscience
- Biomedical Engineering
- Microscopy
Background:
- Miniaturized microscopes allow brain activity visualization in behaving animals.
- Resolving single dendritic spine activity in freely moving subjects remains challenging.
Purpose of the Study:
- To design and apply a fast, high-resolution, miniaturized two-photon microscope (FHIRM-TPM).
- To enable imaging of neuronal activity at the single dendritic spine level in freely behaving animals.
Main Methods:
- Developed a FHIRM-TPM with a lightweight headpiece (2.15 g).
- Utilized a hollow-core photonic crystal fiber for 920-nm femtosecond laser delivery.
- Achieved high spatiotemporal resolution (0.64 μm lateral, 3.35 μm axial) with imaging speeds up to 10,000 Hz (free-line scanning).
Main Results:
- Successfully imaged biosensors (GFP, GCaMP6) at the single dendritic spine level.
- Demonstrated robust, hour-long recordings of neuronal activity in mice during vigorous movements.
- Achieved high spatiotemporal resolution suitable for spine-level analysis.
Conclusions:
- The FHIRM-TPM overcomes previous limitations in imaging neuronal structures in behaving animals.
- This technology facilitates detailed investigation of neural circuit dynamics underlying complex behaviors.
- Enables unprecedented insights into brain function at the single-spine level during naturalistic activity.
