Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy

Lamiae Abdeladim (1) , Katherine S. Matho (1, 2, 3) , Solène Clavreul (2) , Pierre Mahou (1) , Jean-Marc Sintes (1) , Xavier Solinas (1) , Ignacio Arganda-Carreras (4, 5) , Stephen Turney (6) , Jeff Lichtman (6) , Anatole Chessel (1) , Pierre Bemelmans (7) , Karine Loulier (2) , Willy Supatto (1) , Jean Livet (2) , Emmanuel Beaurepaire (1)

1 LOB - Laboratoire d'Optique et Biosciences
2 Institut de la Vision
3 CSHL - Cold Spring Harbor Laboratory
4 UPV/EHU - University of the Basque Country/Euskal Herriko Unibertsitatea
5 Donostia International Physics Center - DIPC (SPAIN)
6 Department of Molecular and Cellular Biology, and Center for Brain Science
7 JACOB - Institut de Biologie François JACOB

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.

Domains: Optics [physics.optics]; Sensory Organs

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