Plastic embedding immunolabeled large-volume samples for three-dimensional high-resolution imaging

塑料嵌入免疫标记大体积样品用于三维高分辨率成像

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作者:Yadong Gang, Xiuli Liu, Xiaojun Wang, Qi Zhang, Hongfu Zhou, Ruixi Chen, Ling Liu, Yao Jia, Fangfang Yin, Gong Rao, Jiadong Chen, Shaoqun Zeng

Abstract

High-resolution three-dimensional biomolecule distribution information of large samples is essential to understanding their biological structure and function. Here, we proposed a method combining large sample resin embedding with iDISCO immunofluorescence staining to acquire the profile of biomolecules with high spatial resolution. We evaluated the compatibility of plastic embedding with an iDISCO staining technique and found that the fluorophores and the neuronal fine structures could be well preserved in the Lowicryl HM20 resin, and that numerous antibodies and fluorescent tracers worked well upon Lowicryl HM20 resin embedding. Further, using fluorescence Micro-Optical sectioning tomography (fMOST) technology combined with ultra-thin slicing and imaging, we were able to image the immunolabeled large-volume tissues with high resolution.

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