Fast scanning high optical invariant two-photon microscopy for monitoring a large neural network activity with cellular resolution

Abstract

Fast and wide imaging with single-cell resolution, high signal-to-noise ratio and no optical aberration has the potential to open up new avenues of investigation in biology. However, this imaging is challenging because of the inevitable tradeoffs among those parameters. Here, we overcome the tradeoffs by combining a resonant scanning system, a large objective with low magnification and high numerical aperture, and highly sensitive large-aperture photodetectors. The result is a practically aberration-free, fast scanning high optical invariant two-photon microscopy (FASHIO-2PM) that enables calcium imaging from a large network composed of ~16k neurons at 7.5 Hz in a 9 mm2 contiguous image plane including more than 10 sensory-motor and higher-order regions of the cerebral cortex in awake mice. Through a network analysis based on single-cell activities, we discover that the brain exhibits small-world-ness rather than scale-freeness. FASHIO-2PM will enable revealing biological dynamics by simultaneous monitoring of macroscopic activity and its composing elements.

Characterizing Cortex-Wide Dynamics with Wide-Field Calcium Imaging

“…Area P has been mainly implicated in visual processing in previous literature (Garrett et al, 2014;Zhuang et al, 2017), and its function in tactile texture discrimination suggests that it may be generally involved in processing information related to object identity (Gilad et al, 2018). With wide-field imaging, these studies provide the first glimpse of the macroscopic activity pattern during sensorimotor Selected references Makino et al, 2017;Musall et al, 2019;Pinto et al, 2019;Peters et al, 2021Bauer et al, 2014Kyriakatos et al, 2017;Kura et al, 2018;Karimi Abadchi et al, 2020Sofroniew et al, 2016Stirman et al, 2016;Ota, 2020;Yu et al, 2020Ji et al, 2016Nöbauer et al, 2017;Song et al, 2017;Weisenburger et al, 2019;Lu et al, 2020Schwalm et al, 2017Schlegel et al, 2018;Jung et al, 2019;Lake et al, 2020 a Within each modality, higher sampling frequency can be achieved at the sacrifice of spatial resolution and vice versa.…”

Fractured columnar small-world functional network organization in volumes of L2/3 of mouse auditory cortex

“…6 A). The small-world propensity can be computed by comparing the clustering coefficient and mean path length of the real data to graphs generated from a Watts–Strogatz model ( 47 , 48 ). To assess small-world propensity, we compared the correlation graph to a regular lattice graph characterized by a high clustering coefficient and a random graph characterized by low mean path length.…”

Expansion-assisted selective plane illumination microscopy for nanoscale imaging of centimeter-scale tissues

“…However, this lens is customized for infrared illumination and exhibits significant field curvature. Additional custom lenses have subsequently been developed for two-photon microscopy, although none of these lenses are suitable for large-scale fluorescence microscopy (Keisuke Ota et al 2020;Rumyantsev et al 2020;Yu et al 2021). A custom lens with a large etendue of G = 18.86 mm 2 , FOV = 14 mm, NA = 0.35, and WD = 19 mm has been developed for the real-time ultra-large-scale high-resolution (RUSH) microscope (Fan et al 2019).…”