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Kilohertz frame-rate two-photon tomography
- Source :
- Nature methods
- Publication Year :
- 2019
-
Abstract
- Point-scanning two-photon microscopy enables high-resolution imaging within scattering specimens such as the mammalian brain, but sequential acquisition of voxels fundamentally limits its speed. We developed a two-photon imaging technique that scans lines of excitation across a focal plane at multiple angles and computationally recovers high-resolution images, attaining voxel rates of over 1 billion Hz in structured samples. Using a static image as a prior for recording neural activity, we imaged visually evoked and spontaneous glutamate release across hundreds of dendritic spines in mice at depths over 250 µm and frame rates over 1 kHz. Dendritic glutamate transients in anesthetized mice are synchronized within spatially contiguous domains spanning tens of micrometers at frequencies ranging from 1–100 Hz. We demonstrate millisecond-resolved recordings of acetylcholine and voltage indicators, three-dimensional single-particle tracking and imaging in densely labeled cortex. Our method surpasses limits on the speed of raster-scanned imaging imposed by fluorescence lifetime. A two-photon computed tomography approach, called scanned line angular projection microscopy, enables high-speed imaging at over 1 kHz frame rates, as demonstrated for glutamate imaging in the in vivo mouse brain.
- Subjects :
- Photon
Glutamic Acid
computer.software_genre
Biochemistry
Article
03 medical and health sciences
Mice
Two-photon excitation microscopy
Voxel
Microscopy
Animals
Molecular Biology
Tomography
030304 developmental biology
Physics
Cerebral Cortex
Neurons
0303 health sciences
Photons
Ranging
Cell Biology
Frame rate
Rats
Mice, Inbred C57BL
Cardinal point
Calcium
Female
computer
Biotechnology
Biomedical engineering
Subjects
Details
- Language :
- English
- ISSN :
- 15487105 and 15487091
- Volume :
- 16
- Issue :
- 8
- Database :
- OpenAIRE
- Journal :
- Nature methods
- Accession number :
- edsair.doi.dedup.....65d9d9d523ced82628b5bbcc41713192