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High-speed neural imaging with multiplexed miniaturized two-photon microscopy.

2025-11-10, Cell reports methods (10.1016/j.crmeth.2025.101221) (online)
Zixiao Zhang, Shing-Jiuan Liu, Ben Mattison, Jessie Muir, Noah Spurr, Christina K Kim, and Weijian Yang (?)
Head-mounted miniaturized two-photon microscopes enable cellular-resolution recording of neural activity deep in the mouse brain during unrestrained behavior. Two-photon microscopy, however, is traditionally limited in frame rate by the necessity of scanning the excitation beam over a large field-of-view (FOV). Here, we present two types of multiplexed miniaturized two-photon microscopes (M-MINI2Ps) that preserve spatial resolution while increasing frame rate by simultaneously imaging two FOVs and demixing them temporally or computationally. We demonstrate large-scale (500 × 500 μm FOV) multiplane calcium imaging in visual and prefrontal cortices of freely moving mice during spontaneous exploration, social behavior, and auditory stimulus. The increased speed of M-MINI2Ps also enables two-photon voltage imaging at 400 Hz over a 380 × 150 μm FOV in freely moving mice. With compact footprints and compatibility with the open-source MINI2P, M-MINI2Ps enable high-speed recording of rapid neural dynamics and large-volume population activity in freely moving mice, providing a powerful tool for systems neuroscience.
This article is included in 1 public curation:

Progress in Voltage Imaging
 
 
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