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Cell-specific three-photon-fluorescence brain imaging: neurons, astrocytes, and gliovascular interfaces

A. A. Lanin, M. S. Pochechuev, A. S. Chebotarev, I. V. Kelmanson, D. S. Bilan, D. A. Kotova, V. S. Tarabykin, A. A. Ivanov, A. B. Fedotov, V. V. Belousov, and A. M. Zheltikov

Abstract

We present brain imaging experiments on rat cortical areas, demonstrating that, when combined with a suitable high-brightness, cell-specific genetically encoded fluorescent marker, three-photon-excited fluorescence (3PEF), enables subcellular-resolution, cell-specific 3D brain imaging that is fully compatible and readily integrable with other nonlinear-optical imaging modalities, including two-photon-fluorescence and harmonic-generation microscopy. With laser excitation provided by sub-100-fs, 1.25-µm laser pulses, cell-specific 3PEF from astrocytes and their processes detected in parallel with a three-photon-resonance-enhanced third harmonic from blood vessels is shown to enable a high-contrast 3D imaging of gliovascular interfaces.

© 2020 Optical Society of America

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Supplementary Material (3)

NameDescription
Visualization 1       Full 3D reconstruction of SypHer3s-expressing neurons from 3PEF images.
Visualization 2       Full 3D reconstructions of SypHer3s-expressing neurons from 2PEF images.
Visualization 3       Full 3D reconstruction of the gliovascular unit from 3PEF/THG images.

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