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Plastic embedding immunolabeled large-volume samples for thre
PRV-CMV-GFP virus and RV-∆G-DsRed virus were used to label subsets of cells in the mouse brain by expressing fluorescent proteins. (From BrainVTA)
The viruses used from BrainVTA in this article are in the table below
RV  RV-∆G-DsRed
Yadong Gang, Xiuli Liu, Xiaojun Wang, Qi Zhang, Hongfu Zhou, Ruixi Chen, Ling Liu, Yao Jia, Fangfang Yin, Gong Rao, Jiadong Chen, Shaoqun Zeng
Pub Date: 2017-09-01, DOI: 10.1364/boe.8.003583, Email: [email protected]
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.

Figure 1. Preservation of immunofluorescence labeled fine details of neurite in resin-embedded brain tissue.
In this study, the authors first examined the proper resin. They found Lowicryl HM20 was a good candidate for this purpose. The fine structures of neurite were preserved in Lowicryl HM20, and that multiple antibodies and fluorescent tracers worked well. They then confirmed that Lowicryl HM20 resin embedding was compatible with the iDISCO staining technique. By combining fMOST technology with resin embedding, they present here an effective approach to visualize molecularly labeled structures in large-volume biological tissue at high resolution.
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