Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy

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.
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Contributor : Aurélien Arnoux <>
Submitted on : Monday, April 29, 2019 - 1:27:39 PM
Last modification on : Monday, July 8, 2019 - 2:59:27 PM


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Lamiae Abdeladim, Katherine Matho, Solène Clavreul, Pierre Mahou, Jean-Marc Sintes, et al.. Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy. Nature Communications, Nature Publishing Group, 2019, 10 (1), pp.1662. ⟨⟩. ⟨10.1038/s41467-019-09552-9⟩. ⟨hal-02114066⟩



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