TY - GEN
T1 - Chromatic serial multiphoton microscopy for multicolor imaging of large brain volumes
AU - Abdeladim, Lamiae
AU - Matho, Katherine S.
AU - Clavreul, Solène
AU - Mahou, Pierre
AU - Sintes, Jean Marc
AU - Solinas, Xavier
AU - Arganda-Carreras, Ignacio
AU - Chessel, Anatole
AU - Turney, Steve
AU - Lichtman, Jeff W.
AU - Bemelmans, Alexis Pierre
AU - Loulier, Karine
AU - Supatto, Willy
AU - Livet, Jean
AU - Beaurepaire, Emmanuel
N1 - Publisher Copyright:
© 2019 SPIE.
PY - 2019/1/1
Y1 - 2019/1/1
N2 - 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.
AB - 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.
KW - Brainbow
KW - Multiphoton microscopy
KW - Neurophotonics
KW - Tissue imaging
U2 - 10.1117/12.2526947
DO - 10.1117/12.2526947
M3 - Conference contribution
AN - SCOPUS:85074251818
T3 - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
BT - Advances in Microscopic Imaging II
A2 - Beaurepaire, Emmanuel
A2 - Pavone, Francesco Saverio
PB - SPIE
T2 - Advances in Microscopic Imaging II 2019
Y2 - 26 June 2019 through 27 June 2019
ER -