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Related Experiment Video

Updated: May 2, 2026

Simultaneous Imaging of Microglial Dynamics and Neuronal Activity in Awake Mice
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High-speed neural imaging with multiplexed miniaturized two-photon microscopy.

Zixiao Zhang1, Shing-Jiuan Liu1, Ben Mattison2

  • 1Department of Electrical and Computer Engineering, University of California, Davis, Davis, CA 95616, USA.

Cell Reports Methods
|November 11, 2025
PubMed
Summary

Multiplexed miniaturized two-photon microscopes (M-MINI2Ps) boost imaging speed for neural activity recording in mice. These advanced microscopes capture rapid neural dynamics and large neural populations in unrestrained subjects.

Keywords:
CP: imagingM-MINI2Pbeam multiplexingcalcium imagingfreely behavingfreely movingminiaturized two-photon microscopetwo-photon miniscopevoltage imaging

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Area of Science:

  • Neuroscience
  • Biomedical Engineering
  • Optical Imaging

Background:

  • Head-mounted miniaturized two-photon microscopes allow cellular-resolution neural recording in awake, behaving mice.
  • Traditional two-photon microscopy is limited by slow scanning speeds, hindering the capture of rapid neural dynamics.

Purpose of the Study:

  • To develop novel multiplexed miniaturized two-photon microscopes (M-MINI2Ps) that enhance imaging frame rates.
  • To enable high-speed, large-volume neural population recording in freely moving mice.

Main Methods:

  • Designed two types of M-MINI2Ps capable of simultaneously imaging two fields-of-view (FOVs).
  • Employed temporal or computational demixing to process data from multiple FOVs.
  • Validated M-MINI2Ps for multiplane calcium imaging and two-photon voltage imaging in mouse cortices.

Main Results:

  • Achieved high-speed, large-scale (500 × 500 μm²) multiplane calcium imaging in visual and prefrontal cortices.
  • Demonstrated two-photon voltage imaging at 400 Hz over a 380 × 150 μm² FOV in freely moving mice.
  • M-MINI2Ps preserved spatial resolution while significantly increasing effective frame rates.

Conclusions:

  • M-MINI2Ps offer a powerful tool for high-speed neural recordings in systems neuroscience.
  • The technology facilitates the study of rapid neural dynamics and large-scale neural population activity in behaving animals.
  • Compact and open-source compatible, M-MINI2Ps enhance accessibility for neuroscience research.