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.
🔬 Techniques
✨ Fluorophores
🧪 Sample Preparation
🏭 Microscope Brands
🔴 Lasers
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Fig. 1
a, Portable two-photon microscope. Electrical lines control the MEMS scanner and focusing micromotor. b, The scanner die is wirebonded onto electrodes on the printed circuit board (PCB). c, Computer-a...
Fig. 2
(Color online) a, Optical deflection angle versus dc voltage, and b, frequency response function for inner (blue solid curve) and outer (red dashed curve) axes of a 1 mm u00d71 mm MEMS scanner. For ea...
Fig. 3
au2013c, Images of neocortical capillaries, averaged over eight frames acquired over 2 s at 4 Hz. du2013f, Line images taken by driving only the scanneru2019s outer axis, at its resonant frequency (56...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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