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Related Experiment Video

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Simultaneous In Vivo Electrophysiology, Two-Photon Imaging, and Optogenetics for Probing Neurovascular Coupling.

Dalchand Ahirwar1, Kun Xie1, Philip O'Herron1

  • 1Department of Physiology, Augusta University, Augusta, GA 30912, USA.

Methods and Protocols
|May 27, 2026
PubMed
Summary

This study introduces a novel method combining electrophysiology, two-photon imaging, and optogenetics to simultaneously measure brain activity and blood flow. This powerful tool advances understanding of neurovascular coupling and its role in neurological disorders.

Keywords:
electrophysiologyfunctional hyperemialocal field potentialmultimodal imagingneural dynamicsneurovascular couplingtwo-photon imaging

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Area of Science:

  • Neuroscience
  • Vascular Biology
  • Systems Biology

Background:

  • Neurovascular coupling, the link between neuronal activity and cerebral blood flow, is crucial for brain metabolism.
  • Its disruption is implicated in neurodegenerative diseases like Alzheimer's, Parkinson's, stroke, and aging.
  • Understanding these mechanisms requires simultaneous high-resolution measurement of neural and vascular dynamics with perturbation capabilities.

Purpose of the Study:

  • To present a novel methodological framework for in vivo investigation of neurovascular coupling.
  • To enable simultaneous measurement of neuronal activity, cerebral blood flow, and vascular dynamics.
  • To allow targeted optogenetic manipulation of the vasculature during neural and vascular recordings.

Main Methods:

  • Chronic electrophysiological recordings using flexible electrode arrays for single-unit activity, multi-unit activity, and local field potentials.
  • Two-photon microscopy for high-resolution imaging of cerebral blood flow and individual vessel diameter.
  • Optogenetic manipulation of vascular smooth muscle cells for rapid, reversible control of vessel diameter.

Main Results:

  • Demonstration of a combined platform for simultaneous, high-resolution recording of neural and vascular activity in vivo.
  • Successful optogenetic manipulation of vascular tone while concurrently monitoring neural and vascular dynamics.
  • Detailed protocols for implantation, data acquisition, and analysis are provided.

Conclusions:

  • The presented combined platform is a powerful tool for mechanistic studies of neurovascular coupling.
  • This methodology facilitates research into the dysfunction of neurovascular coupling in various disease models.
  • It offers unprecedented capabilities for investigating the intricate relationship between brain activity and blood flow.