Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy

Lamiae Abdeladim, Katherine S. Matho, Solène Clavreul, Pierre Mahou, Jean-Marc Sintes, Xavier Solinas, Ignacio Arganda-Carreras, Stephen G. Turney, Jeff W. Lichtman, Anatole Chessel, Alexis-Pierre Bemelmans, Karine Loulier, Willy Supatto, Jean Livet () and Emmanuel Beaurepaire ()
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Lamiae Abdeladim: Ecole polytechnique, CNRS, INSERM, IP Paris
Katherine S. Matho: Ecole polytechnique, CNRS, INSERM, IP Paris
Solène Clavreul: CNRS, Institut de la Vision
Pierre Mahou: Ecole polytechnique, CNRS, INSERM, IP Paris
Jean-Marc Sintes: Ecole polytechnique, CNRS, INSERM, IP Paris
Xavier Solinas: Ecole polytechnique, CNRS, INSERM, IP Paris
Ignacio Arganda-Carreras: University of the Basque Country
Stephen G. Turney: Harvard University
Jeff W. Lichtman: Harvard University
Anatole Chessel: Ecole polytechnique, CNRS, INSERM, IP Paris
Alexis-Pierre Bemelmans: Université Paris-Sud
Karine Loulier: CNRS, Institut de la Vision
Willy Supatto: Ecole polytechnique, CNRS, INSERM, IP Paris
Jean Livet: CNRS, Institut de la Vision
Emmanuel Beaurepaire: Ecole polytechnique, CNRS, INSERM, IP Paris

Nature Communications, 2019, vol. 10, issue 1, 1-14

Abstract: Abstract Large-scale microscopy approaches are transforming brain imaging, but currently lack efficient multicolor contrast modalities. We introduce chromatic multiphoton serial (ChroMS) microscopy, a method integrating one‐shot multicolor multiphoton excitation through wavelength mixing and serial block-face image acquisition. This approach provides organ-scale micrometric imaging of spectrally distinct fluorescent proteins and label-free nonlinear signals with constant micrometer-scale resolution and sub-micron channel registration over the entire imaged volume. We demonstrate tridimensional (3D) multicolor imaging over several cubic millimeters as well as brain-wide serial 2D multichannel imaging. We illustrate the strengths of this method through color-based 3D analysis of astrocyte morphology and contacts in the mouse cerebral cortex, tracing of individual pyramidal neurons within densely Brainbow-labeled tissue, and multiplexed whole-brain mapping of axonal projections labeled with spectrally distinct tracers. ChroMS will be an asset for multiscale and system-level studies in neuroscience and beyond.

Date: 2019
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DOI: 10.1038/s41467-019-09552-9

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