Chromatic serial multiphoton microscopy for multicolor imaging of large brain volumes

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

doi:10.1117/12.2526947

22 July 2019 Chromatic serial multiphoton microscopy for multicolor imaging of large brain volumes

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

Author Affiliations +

Proceedings Volume 11076, Advances in Microscopic Imaging II; 1107607 (2019) https://doi.org/10.1117/12.2526947
Event: European Conferences on Biomedical Optics, 2019, Munich, Germany

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.

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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, and Emmanuel Beaurepaire "Chromatic serial multiphoton microscopy for multicolor imaging of large brain volumes", Proc. SPIE 11076, Advances in Microscopic Imaging II, 1107607 (22 July 2019); https://doi.org/10.1117/12.2526947

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KEYWORDS

Multiphoton microscopy

Brain

Microscopy

Neuroimaging

Colorimetry

Tissues

Image resolution

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