The emergence of various optical clearing methods provides a great potential for imaging deep inside tissues by combining with multiple-labelling and microscopic imaging techniques. They were generally developed for specific imaging demand thus presented some non-negligible limitations such as long incubation time, tissue deformation, fluorescence quenching, incompatibility with immunostaining or lipophilic tracers. In this study, we developed a rapid and versatile clearing method, termed ReagentTF, for deep imaging of various fluorescent samples. This method can not only efficiently clear embryos, neonatal whole-brains and adult thick brain sections by simple immersion in aqueous mixtures with minimal volume change, but also can preserve fluorescence of various fluorescent proteins and simultaneously be compatible with immunostaining and lipophilic neuronal dyes. We demonstrate the effectiveness of this method in reconstructing the cell distributions of mouse hippocampus, visualizing the neural projection from CA1 (Cornu Ammonis 1) to HDB (nucleus of the horizontal limb of the diagonal band), and observing the growth of forelimb plexus in whole-mount embryos. These results suggest that ReagentTF is useful for large-volume imaging and will be an option for the deep imaging of biological tissues.
Tingting Yu, Jingtan Zhu, Yusha Li, Yisong Qi, Jianyi Xu, Hui Gong, Qingming Luo, and Dan Zhu, "ReagentTF: a rapid and versatile optical clearing method for biological imaging (Conference Presentation)," Proc. SPIE 10051, Neural Imaging and Sensing, 100510J (Presented at SPIE BiOS: January 31, 2017; Published: 19 April 2017); https://doi.org/10.1117/12.2253491.5371358594001.
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