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High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

Author

Listed:
  • Jiang Lan Fan

    (University of California
    University of California)

  • Jose A. Rivera

    (University of California)

  • Wei Sun

    (Thorlabs Imaging Systems)

  • John Peterson

    (Thorlabs Imaging Systems)

  • Henry Haeberle

    (Thorlabs Imaging Systems)

  • Sam Rubin

    (Thorlabs Imaging Systems
    LightPath Technologies Inc.)

  • Na Ji

    (University of California
    University of California
    University of California
    Lawrence Berkeley National Laboratory)

Abstract

Understanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

Suggested Citation

  • Jiang Lan Fan & Jose A. Rivera & Wei Sun & John Peterson & Henry Haeberle & Sam Rubin & Na Ji, 2020. "High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics," Nature Communications, Nature, vol. 11(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-19851-1
    DOI: 10.1038/s41467-020-19851-1
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