Open AccessPlastic embedding immunolabeled large-volume samples for three-dimensional high-resolution imaging
Author(s) -
Yadong Gang,
Xiuli Liu,
Xiaojun Wang,
Qi Zhang,
Haiyan Zhou,
Ruixi Chen,
Ling Liu,
Jia Yao,
FangFang Yin,
Gang Rao,
Jiadong Chen,
Shaoqun Zeng
Publication year - 2017
Publication title -
biomedical optics express
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.362
H-Index - 86
ISSN - 2156-7085
DOI - 10.1364/boe.8.003583
Subject(s) - materials science , fluorescence , biomolecule , resolution (logic) , biological specimen , high resolution , image resolution , biomedical engineering , slicing , embedding , microscopy , optics , nanotechnology , computer science , artificial intelligence , physics , medicine , remote sensing , geology , world wide web
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.