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Proceedings Paper

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, Lab for Optics and Biosciences, CNRS, INSERM (France)
Katherine S. Matho, Ecole Polytechnique, Lab for Optics and Biosciences, CNRS, INSERM (France)
Sorbonne Univ., Institut de la Vision (France)
CSHL (United States)
Solène Clavreul, Sorbonne Univ., INSERM, CNRS, Institut de la Vision (France)
Pierre Mahou, Ecole Polytechnique, Lab for Optics and Biosciences, CNRS, INSERM (France)
Jean-Marc Sintes, Ecole Polytechnique, Lab for Optics and Biosciences, CNRS, INSERM (France)
Xavier Solinas, Ecole Polytechnique, Lab for Optics and Biosciences, CNRS, INSERM (France)
Ignacio Arganda-Carreras, IKERBASQUE (Spain)
Anatole Chessel, Ecole Polytechnique, Lab for Optics and Biosciences, 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, Institut de la Vision (France)
Willy Supatto, Ecole Polytechnique, Lab for Optics and Biosciences, CNRS, INSERM (France)
Jean Livet, Sorbonne Univ., INSERM, CNRS, Institut de la Vision (France)
Emmanuel Beaurepaire, Ecole Polytechnique, Lab for Optics and Biosciences, 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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