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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.

Author information

1
Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
2
MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.

Abstract

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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