Journal
NATURE NEUROSCIENCE
Volume 16, Issue 8, Pages 1154-U246Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nn.3447
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Funding
- PRESTO program of the Japan Science and Technology Agency
- Sumitomo Foundation
- Nakajima Foundation
- Mitsubishi Foundation
- Strategic Programs for R&D (President's Discretionary Fund) of RIKEN
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- Grants-in-Aid for Scientific Research [24650175, 23680038] Funding Source: KAKEN
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We report a water-based optical clearing agent, SeeDB, which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. Our method maintained a constant sample volume during the clearing procedure, an important factor for keeping cellular morphology intact, and facilitated the quantitative reconstruction of neuronal circuits. Combined with two-photon microscopy and an optimized objective lens, we were able to image the mouse brain from the dorsal to the ventral side. We used SeeDB to describe the near-complete wiring diagram of sister mitral cells associated with a common glomerulus in the mouse olfactory bulb. We found the diversity of dendrite wiring patterns among sister mitral cells, and our results provide an anatomical basis for non-redundant odor coding by these neurons. Our simple and efficient method is useful for imaging intact morphological architecture at large scales in both the adult and developing brains.
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