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Adapting a Two-Photon Scanning Microscope for Simultaneous Single-Photon Imaging of an Infrared Dopamine Sensor.

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  • 1Department of Neuroscience and Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, California 94720.

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Summary

We developed a new method to upgrade two-photon microscopes for simultaneous visible and near-infrared fluorescence imaging. This technique enhances optical sectioning and reduces background noise for improved neuroscience research and dopamine sensor comparisons.

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Area of Science:

  • Neuroscience
  • Biomedical Imaging
  • Microscopy

Background:

  • Simultaneous dual-color imaging is crucial for neuroscience and biomedical research.
  • Two-photon microscopy is a standard technique, but lacks efficient near-infrared detection.
  • Near-infrared wavelengths minimize scattering and autofluorescence in biological samples.

Purpose of the Study:

  • To adapt a two-photon scanning microscope for simultaneous detection of visible and near-infrared fluorescence.
  • To validate the new method by comparing dopamine sensors and imaging neural activity.

Main Methods:

  • Modified a two-photon scanning microscope to include a near-infrared detection channel using a single-mode optical fiber.
  • Used the modified microscope to compare genetically encoded dopamine sensors (GRABDA) and carbon nanotube-based sensors (nIRCats).
  • Simultaneously imaged calcium changes and dopamine release in mouse retina.

Main Results:

  • The fiber coupling improved optical sectioning and reduced background noise for near-infrared signals.
  • Demonstrated the first direct comparison between GRABDA and nIRCats dopamine sensors.
  • Successfully performed simultaneous imaging of calcium activity and dopamine release in the retina.

Conclusions:

  • The adapted two-photon microscope enables simultaneous functional imaging in visible and near-infrared channels.
  • This platform is versatile and applicable to various fluorophores and existing two-photon microscopes.
  • The method advances dual-color imaging capabilities in neuroscience and biomedical research.