Video-rate volumetric functional imaging of the brain at synaptic resolution
Rongwen Lu,Wenzhi Sun,Yajie Liang,Aaron M. Kerlin,Jens Bierfeld,Johannes D. Seelig,Daniel E. Wilson,Benjamin Scholl,Boaz Mohar,Masashi Tanimoto,Masashi Tanimoto,Minoru Koyama,David Fitzpatrick,Michael B. Orger,Na Ji +14 more
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TLDR
An optical module is presented that is easily integrated into standard two-photon laser-scanning microscopes to generate an axially elongated Bessel focus, which when scanned in 2D turns frame rate into volume rate.Abstract:
Neurons and neural networks often extend hundreds of micrometers in three dimensions. Capturing the calcium transients associated with their activity requires volume imaging methods with subsecond temporal resolution. Such speed is a challenge for conventional two-photon laser-scanning microscopy, because it depends on serial focal scanning in 3D and indicators with limited brightness. Here we present an optical module that is easily integrated into standard two-photon laser-scanning microscopes to generate an axially elongated Bessel focus, which when scanned in 2D turns frame rate into volume rate. We demonstrated the power of this approach in enabling discoveries for neurobiology by imaging the calcium dynamics of volumes of neurons and synapses in fruit flies, zebrafish larvae, mice and ferrets in vivo. Calcium signals in objects as small as dendritic spines could be resolved at video rates, provided that the samples were sparsely labeled to limit overlap in their axially projected images.read more
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Neuronal cell-type classification: challenges, opportunities and the path forward
Hongkui Zeng,Joshua R. Sanes +1 more
TL;DR: In this paper, a staged approach for cell type classification in the brain is proposed, including the incorporation of multiple, quantitative features as criteria, the use of discontinuous variation to define types and the creation of a hierarchical system to represent relationships between cells.
Journal ArticleDOI
Genetic Dissection of Neural Circuits: A Decade of Progress.
TL;DR: Evaluated methods allow systematic classification of cell types and provide genetic access to diverse neuronal types for studies of connectivity and neural coding during behavior in invertebrate and vertebrate organisms.
Journal ArticleDOI
In vivo imaging of neural activity
Weijian Yang,Rafael Yuste +1 more
TL;DR: Advanced microscopy techniques for in vivo functional imaging are reviewed and guidelines for which technologies are best suited for particular applications are offered.
Journal ArticleDOI
High-speed volumetric imaging of neuronal activity in freely moving rodents.
Oliver Skocek,Tobias Nöbauer,Lukas Weilguny,Francisca Martínez Traub,Chuying Naomi Xia,Maxim I. Molodtsov,Abhinav Grama,Masahito Yamagata,Daniel Aharoni,David D. Cox,Peyman Golshani,Peyman Golshani,Alipasha Vaziri,Alipasha Vaziri +13 more
TL;DR: A miniature light-field microscope in combination with a signal-extraction approach enables high-speed volumetric calcium imaging in freely moving mice and it is demonstrated that neurons separated by as little as ~15 µm and at depths up to 360‵m can be discriminated.
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