Journal
CELL
Volume 157, Issue 3, Pages 726-739Publisher
CELL PRESS
DOI: 10.1016/j.cell.2014.03.042
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Funding
- Program for Innovative Cell Biology by Innovative Technology
- Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan
- MEXT/Japan Society for the Promotion of Science (JSPS) [25221004, 23115006]
- strategic programs for R&D (President's discretionary fund) of RIKEN
- RIKEN Center for Developmental Biology and RIKEN Quantitative Biology Center
- Core Research for Evolutional Science and Technology (CREST)
- Precursory Research for Embryonic Science and Technology (PRESTO) of Japan Science and Technology Agency (JST)
- RIKEN Special Postdoctoral Research Program
- Japan Foundation for Applied Enzymology
- Shimabara Science Promotion Foundation
- RIKEN Foreign Postdoctoral Researcher Program
- Grants-in-Aid for Scientific Research [25560428, 26280110, 26710014, 26640134] Funding Source: KAKEN
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Systems-level identification and analysis of cellular circuits in the brain will require the development of whole-brain imaging with single-cell resolution. To this end, we performed comprehensive chemical screening to develop a whole-brain clearing and imaging method, termed CUBIC (clear, unobstructed brain imaging cocktails and computational analysis). CUBIC is a simple and efficient method involving the immersion of brain samples in chemical mixtures containing aminoalcohols, which enables rapid whole-brain imaging with single-photon excitation microscopy. CUBIC is applicable to multicolor imaging of fluorescent proteins or immunostained samples in adult brains and is scalable from a primate brain to subcellular structures. We also developed a whole-brain cell-nuclear counterstaining protocol and a computational image analysis pipeline that, together with CUBIC reagents, enable the visualization and quantification of neural activities induced by environmental stimulation. CUBIC enables time-course expression profiling of whole adult brains with single-cell resolution.
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