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

Optics Letters
  • Alexandr Lanin, Matvey Pochechuev, artem chebotarev, ilya kelmanson, Dmitry Bilan, Darya Kotova, Victor Tarabykin, Anatoly Ivanov, Andrey Fedotov, Vsevolod Belousov, and Aleksei Zheltikov
  • received 09/11/2019; accepted 11/04/2019; posted 11/05/2019; Doc. ID 377494
  • 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 modality 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.
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