Scalable and DiI-compatible optical clearance of the mammalian brain

Frontiers in Neuroanatomy

Zaifu eYang¹  Bing eHou¹  Shaoxia eWang¹  Jiarui eWang¹  Yang eLi¹  Dan eZhang²  Lingzhong eFan²  Tianzi eJiang²  Bing eLiu²  Shan eZhao²  Xin eZhang²  Zilong eQiu³  Mengping eWei⁴  Chen eZhang⁴ 

[1] Beijing Institute of Radiation Medicine;Institute of Automation, Chinese Academy of Sciences;Institute of Neuroscience, Shanghai Institutes for Biological Sciences;Peking University;School of Life Science and Technology, University of Electronic Science and Technology of China;The University of Queensland;

关键词: Urea;    whole brain imaging;    tract tracing;    CLARITY;    Cubic;    Optical tissue clearing;   

DOI  :  10.3389/fnana.2015.00019

来源: DOAJ

【 摘 要 】

Efficient optical clearance is fundamental for whole brain imaging. In particular, clearance of the brain without membrane damage is required for the imaging of lipophilic tracer-labeled neural tracts. Relying on an ascending gradient of fructose solutions, SeeDB can achieve sufficient transparency of the mouse brain while ensuring that the plasma membrane remains intact. However, it is challenging to extend this method to larger mammalian brains due to the extremely high viscosity of the saturated fructose solution. Here we report a SeeDB-derived optical clearing method, termed FRUIT, which utilizes a cocktail of fructose and urea. As demonstrated in the adult mouse brain, combination of these two highly water-soluble clearing agents exerts a synergistic effect on clearance. More importantly, the final FRUIT solution has low viscosity so as to produce transparency of the whole adult rabbit brain via arterial perfusion, which is impossible to achieve with a saturated fructose solution. In addition to good compatibility with enhanced yellow fluorescent protein, the cocktail also preserves the fluorescence of the lipophilic tracer DiI. This work provides a volume-independent optical clearing method which retains the advantages of SeeDB, particularly compatibility with lipophilic tracers.

【 授权许可】

Unknown