Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Dual-plane 3-photon microscopy with remote focusing.

Kevin T Takasaki1, Dmitri Tsyboulski2, Jack Waters1

  • 1Allen Institute for Brain Science, 615 Westlake Ave N, Seattle, WA 98109, USA.

Biomedical Optics Express
|December 5, 2019
PubMed
Summary

Dual-plane 3-photon microscopy doubles imaging speed deep in scattering tissues. This advancement enables faster in vivo calcium imaging in dense samples, overcoming previous limitations.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Multimodal characterization of variation in neuronal types in the mouse basal ganglia.

bioRxiv : the preprint server for biology·2026
Same author

Connecting single-cell transcriptomes to projectomes in the mouse visual cortex.

Nature·2026
Same author

Morphoelectric Diversity and Specialization of Neuronal Cell Types in the Primate Striatum.

bioRxiv : the preprint server for biology·2026
Same author

A Cross-Species Enhancer-AAV Toolkit for Cell Type-Specific Targeting Across the Basal Ganglia.

bioRxiv : the preprint server for biology·2026
Same author

<i>MapMyCells:</i> High-performance mapping of unlabeled cell-by-gene data to reference brain taxonomies.

bioRxiv : the preprint server for biology·2026
Same author

A consensus spinal cord cell type atlas across mouse, macaque, and human.

bioRxiv : the preprint server for biology·2026

Area of Science:

  • Neuroscience
  • Biomedical Optics
  • Microscopy

Background:

  • 3-photon excitation microscopy allows deep tissue imaging in scattering samples.
  • Current 3-photon microscopy is limited to single-focus scanning, restricting volumetric imaging speed.

Purpose of the Study:

  • To implement and characterize dual-plane 3-photon microscopy for enhanced in vivo imaging.
  • To demonstrate simultaneous calcium imaging in two distinct planes deep within the brain.

Main Methods:

  • Developed a dual-plane 3-photon microscopy system using temporal multiplexing and remote focusing.
  • Performed simultaneous in vivo calcium imaging in mouse cortex at depths exceeding 600 µm.
  • Utilized pan-excitatory GCaMP6s transgenic mice for fluorescence detection.

Related Experiment Videos

Main Results:

  • Achieved simultaneous imaging of two planes with a per-plane framerate of 7 Hz.
  • Demonstrated effective 2 MHz laser repetition rate for rapid data acquisition.
  • Validated the method's applicability for deep cortical imaging in scattering biological tissue.

Conclusions:

  • Dual-plane 3-photon microscopy significantly doubles volumetric imaging rates compared to single-plane systems.
  • This technique is a generalizable modification for existing 3-photon excitation (3PE) systems.
  • The method uses off-the-shelf components, offering a practical approach for accelerated deep-tissue neuroscience research.