Article Synopsis
- Researchers developed a lightweight three-photon microscope (m3PM) that improves fluorescence collection for deep-tissue imaging, especially in highly scattering environments.
- The m3PM is capable of imaging calcium activity in the cortex and dorsal hippocampal CA1 up to 1.2 mm deep while using safe laser power.
- It can also detect neuron activities related to sensorimotor behavior in freely moving mice during specific tasks, enabling deeper insights into neural mechanisms in various brain regions.
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Article Abstract
In deep-tissue multiphoton microscopy, diffusion and scattering of fluorescent photons, rather than ballistic emanation from the focal point, have been a confounding factor. Here we report on a 2.17-g miniature three-photon microscope (m3PM) with a configuration that maximizes fluorescence collection when imaging in highly scattering regimes. We demonstrate its capability by imaging calcium activity throughout the entire cortex and dorsal hippocampal CA1, up to 1.2 mm depth, at a safe laser power. It also enables the detection of sensorimotor behavior-correlated activities of layer 6 neurons in the posterior parietal cortex in freely moving mice during single-pellet reaching tasks. Thus, m3PM-empowered imaging allows the study of neural mechanisms in deep cortex and subcortical structures, like the dorsal hippocampus and dorsal striatum, in freely behaving animals.
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