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Biomed Opt Express. 2019 Apr 29;10(5):2612-2622. doi: 10.1364/BOE.10.002612. eCollection 2019 May 1.

On-line optical clearing method for whole-brain imaging in mice.

Wu H1,2, Yang X1,2, Chen S1,2, Zhang L1,2, Long B1,2, Tan C1,2, Yuan J1,2, Gong H1,2.

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Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.


The combination of optical clearing with light microscopy has a number of applications in the whole-brain imaging of mice. However, the initial processing time of optical clearing is time consuming, and the protocol is complicated. We propose a novel method based on on-line optical clearing. Agarose-embedded mouse brain was immersed in the optical clearing reagent, and clearing of the brain was achieved ~100 μm beneath the sample surface. After imaging, the cleared layer was removed, thereby allowing layer-by-layer clearing and imaging. No pre-immersion was required, and we demonstrated that on-line optical clearing can reduce the whole-brain imaging time by half.

Conflict of interest statement

The authors declare that there are no conflicts of interest related to this article.

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