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Multiphoton Intravital Calcium Imaging.

Claire E J Cheetham1

  • 1Department of Neurobiology, University of Pittsburgh, Pittsburgh, Pennsylvania.

Current Protocols in Cytometry
|June 27, 2018
PubMed
Summary
This summary is machine-generated.

Multiphoton intravital calcium imaging allows high-resolution monitoring of cellular activity deep within living organisms. This study details protocols for two-photon microscopy of genetically encoded calcium indicators in mouse brains.

Keywords:
2-photon microscopybraincranial windowgenetically-encoded calcium indicatorin vivo imagingintravital imagingmultiphoton imaging

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Area of Science:

  • Neuroscience
  • Biomedical Engineering
  • Microscopy

Background:

  • Multiphoton intravital calcium imaging offers high-resolution, longitudinal monitoring of cellular activity deep within living organisms.
  • Genetically encoded calcium indicators (GECIs) combined with two-photon microscopy enable visualization of neural activity.
  • Real-time imaging of intracellular calcium concentration in the mouse brain is crucial for understanding neural circuit function.

Purpose of the Study:

  • To present protocols for applying two-photon microscopy to image GECIs in the mouse brain.
  • To enable real-time, high-resolution imaging of neuronal activity and synaptic function in vivo.
  • To provide methods for cranial window implantation and image acquisition for longitudinal studies.

Main Methods:

  • Two-photon microscopy for imaging genetically encoded calcium indicators (GECIs) in the mouse brain.
  • Surgical implantation of cranial imaging windows (open skull and thinned skull) for optical access.
  • Protocols for image acquisition during sensory stimulation or behavioral tasks.

Main Results:

  • Enables simultaneous imaging of intracellular calcium concentration in hundreds of neurons.
  • Achieves imaging resolution down to the level of single synapses.
  • Provides methods for both single and multi-session imaging.

Conclusions:

  • This unit provides essential protocols for advanced in vivo neural imaging in the mouse brain.
  • The described techniques facilitate the study of neural dynamics during behavior and sensory processing.
  • The methods support high-resolution, longitudinal monitoring of cellular and subcellular activity in neuroscience research.