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

Jiang Lan Fan, Jose A. Rivera, Wei Sun, John Peterson, Henry Haeberle, Sam Rubin and Na Ji ()
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Jiang Lan Fan: 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
Na Ji: University of California

Nature Communications, 2020, vol. 11, issue 1, 1-12

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-19851-1

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