Miniature three-photon microscopy maximized for scattered fluorescence collection.
Chunzhu ZhaoShiyuan ChenLifeng ZhangDong ZhangRunlong WuYanhui HuFengqingyang ZengYijun LiDakun WuFei YuYunfeng ZhangJue ZhangLiangyi ChenAimin WangHeping Peace ChengPublished in: Nature methods (2023)
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.
Keyphrases
- high resolution
- spinal cord
- single molecule
- functional connectivity
- neuropathic pain
- living cells
- label free
- optical coherence tomography
- high speed
- particulate matter
- working memory
- high throughput
- heavy metals
- mass spectrometry
- risk assessment
- white matter
- adipose tissue
- fluorescence imaging
- blood brain barrier
- energy transfer