Multicolor multiscale brain imaging with chromatic multiphoton serial microscopy

Nat Commun. 2019 Apr 10;10(1):1662. doi: 10.1038/s41467-019-09552-9.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Astrocytes / metabolism
  • Cerebral Cortex / cytology
  • Cerebral Cortex / diagnostic imaging*
  • Color
  • Dependovirus
  • Female
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • HEK293 Cells
  • Humans
  • Imaging, Three-Dimensional / methods*
  • Luminescent Proteins / chemistry*
  • Luminescent Proteins / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Microscopy, Fluorescence, Multiphoton / methods*
  • Models, Animal
  • Nestin / genetics
  • Neuroanatomical Tract-Tracing Techniques / methods
  • Neuroimaging / methods*
  • Parvovirinae / genetics
  • Pyramidal Cells / metabolism
  • Transfection

Substances

  • Luminescent Proteins
  • Nes protein, mouse
  • Nestin

Supplementary concepts

  • Adeno-associated virus-2