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

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 similar to 100 mu m beneath the sample surface. After imaging, the cleared layer was removed, thereby allowing layer-bylayer clearing and imaging. No pm-immersion was required, and we demonstrated that online optical clearing can reduce the whole-brain imaging time by half. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement