Murakami TC, Mano T, Saikawa S, Horiguchi SA, Shigeta D, Baba K, Sekiya H, Shimizu Y, Tanaka KF, Kiyonari H, Iino M, Mochizuki H, Tainaka K, Ueda HR. A three-dimensional single-cell-resolution whole-brain atlas using CUBIC-X expansion microscopy and tissue clearing. Nat Neurosci. 2018 Mar 5; PubMed.
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Paris-Saclay University
This new technique allows the three-dimensional imaging of a whole mouse brain with a single-cell resolution without affecting the integrity of the brain analyzed. Brain organization and connections are maintained, and the analysis at such a powerful resolution does not require sectioning and analysis of slices by sampling methods, allowing unbiased quantifications of cellular populations in the entire brain without requiring time-consuming and tedious analysis by unbiased stereology.
The technique developed and the analyses performed here are truly impressive, culminating in a three-dimensional atlas of the mouse brain with a level of detail and precision that could not be achieved with classical techniques, nor with previous clearing methods.
The technique is proven to be efficient when using fluorescence reporter proteins, but ultimately being able to apply it as well to brains stained with classical immunofluorescence techniques will broaden its applications.
In addition, the current whole-brain analysis of the cell population is based on a nuclear marker that is easier to identify than a marker of the cell body. Hopefully, this technique can be expanded to perform single-cell detection of cellular makers localized within the cytoplasm (more challenging due to the diversity of cell sizes and shapes) that would allow cell quantification of specific cellular populations, or even to track single neuron projections throughout the brain. These techniques would allow faster and easier analysis of cell loss in specific cell populations in the whole brain in neurodegenerative models (but not only) and might help us identify more precisely the extent of axonal degeneration, pathological conditions, and the specific pathways involved in disease models.
View all comments by Nolwen ReyMRC Laboratory of Molecular Biology
This paper used whole organ clearing and homogeneous tissue expansion for producing a three-dimensional atlas of the mammalian brain. Nerve cell numbers could be quantitated over time in a given brain region in an unbiased manner. Thus, the authors showed a reduction in nerve cell numbers in the somatosensory cortex and the visual area during early postnatal development. Models of human neurodegenerative diseases often develop filamentous inclusions, but not much is known about the death of cells. We know even less about the effects of seeded, prion-like aggregates on neurodegeneration. Work such as that described here may lend itself to address these important questions.
View all comments by Michel GoedertMake a Comment
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