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

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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