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Chromatic serial multiphoton microscopy for multicolor imaging of large brain volumes
Author(s): Lamiae Abdeladim; Katherine S. Matho; Solène Clavreul; Pierre Mahou; Jean-Marc Sintes; Xavier Solinas; Ignacio Arganda-Carreras; Anatole Chessel; Steve Turney; Jeff W. Lichtman; Alexis-Pierre Bemelmans; Karine Loulier; Willy Supatto; Jean Livet; Emmanuel Beaurepaire
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Paper Abstract

Large-scale microscopy approaches are transforming brain imaging, but currently lack efficient multicolor contrast modalities. We address this issue by introducing chromatic multiphoton serial (ChroMS) microscopy, a method combining multicolor multiphoton excitation through wavelength mixing and microtome-assisted serial block-face image acquisition. This approach delivers large-scale micrometric imaging of spectrally distinct fluorescent proteins with constant micrometer-scale resolution and sub-micron channel registration over the entire imaged volume. We achieve multicolor 3D imaging over several cubic millimeters and brain-wide serial 2D multicolor imaging. We illustrate the potential of this method for several novel types of measurements interesting for region-scale or whole brain studies: (i) color-based analysis of astrocyte morphology and spatial interactions in the mouse cerebral cortex, (ii) tracing of densely labeled neurons, and (iii) brain-wide mapping of axonal projections labeled with distinct tracers.

Paper Details

Date Published: 22 July 2019
PDF: 3 pages
Proc. SPIE 11076, Advances in Microscopic Imaging II, 1107607 (22 July 2019); doi: 10.1117/12.2526947
Show Author Affiliations
Lamiae Abdeladim, Ecole Polytechnique, CNRS, INSERM (France)
Katherine S. Matho, Ecole Polytechnique, CNRS, INSERM (France)
Sorbonne Univ. (France)
CSHL (United States)
Solène Clavreul, Sorbonne Univ., INSERM, CNRS (France)
Pierre Mahou, Ecole Polytechnique, CNRS, INSERM (France)
Jean-Marc Sintes, Ecole Polytechnique, CNRS, INSERM (France)
Xavier Solinas, Ecole Polytechnique, CNRS, INSERM (France)
Ignacio Arganda-Carreras, IKERBASQUE (Spain)
Anatole Chessel, Ecole Polytechnique, CNRS, INSERM (France)
Steve Turney, Harvard Univ. (United States)
Jeff W. Lichtman, Harvard Univ. (United States)
Alexis-Pierre Bemelmans, MIRCen, CEA, CNRS (France)
Karine Loulier, Sorbonne Univ., INSERM, CNRS (France)
Willy Supatto, Ecole Polytechnique, CNRS, INSERM (France)
Jean Livet, Sorbonne Univ., INSERM, CNRS (France)
Emmanuel Beaurepaire, Ecole Polytechnique, CNRS, INSERM (France)

Published in SPIE Proceedings Vol. 11076:
Advances in Microscopic Imaging II
Emmanuel Beaurepaire; Francesco Saverio Pavone; Peter T. C. So, Editor(s)

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