Bessel foci used for two-photon fluorescence excitation have enabled high-speed volumetric imaging. At high numeric aperture, their imaging performances are compromised by substantial side-ring excitation and suffer from reduced image contrast. Here, we describe axially extended Bessel-droplet foci with suppressed side rings and more resistance to optical aberrations. Applying novel phase patterns to generate Bessel-droplet foci of variable NAs at high power throughput, we achieved continuous volume imaging by scanning them interferometrically along the axial direction. With Bessel-droplet foci, we demonstrated high-resolution volumetric imaging of synaptic anatomy and function as well as lymphatic circulations in the mouse brain in vivo.
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Wei Chen, Qinrong Zhang, Ryan Gregory Natan, Jianglan Fan, Na Ji, "High-contrast volumetric imaging of synapses and microcirculations with Bessel-droplet two-photon fluorescence microscopy," Proc. SPIE PC11946, Neural Imaging and Sensing 2022, PC119460N (28 April 2022); https://doi.org/10.1117/12.2610205