Scalable Resin Embedding Method for Large-Volume Brain Tissues with High Fluorescence Preservation Capacity

Ting Luo; Lei Deng; Anan Li; Can Zhou; Shuai Shao; Qingtao Sun; Hui Gong; Xiaoquan Yang; Xiangning Li

doi:10.1016/j.isci.2020.101717

Scalable Resin Embedding Method for Large-Volume Brain Tissues with High Fluorescence Preservation Capacity

iScience. 2020 Oct 20;23(11):101717. doi: 10.1016/j.isci.2020.101717. eCollection 2020 Nov 20.

Authors

Ting Luo^{1

2}, Lei Deng^{1

2}, Anan Li^{1

2

3}, Can Zhou^{1

2}, Shuai Shao^{1

2}, Qingtao Sun³, Hui Gong^{1

2

3}, Xiaoquan Yang^{1

2

3}, Xiangning Li^{1

2

3}

Affiliations

¹ Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, #1037, Luoyu Road, Wuhan, Hubei 430074, China.
² MoE Key Laboratory for Biomedical Photonics, School of Engineering Sciences, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
³ HUST-Suzhou Institute for Brainsmatics, Suzhou 215125, China.

Abstract

Resin embedding is widely used to dissect the fine structure of bio-tissue with electron and optical microscopy. However, it is difficult to embed large-volume tissues with resin. Here, we modified the formula of LR-White resin to prevent the sample cracking during polymerization process and applied this method to the intact brains of mouse, ferret, and macaque. Meanwhile, we increased the fluorescence preservation rate for green fluorescent protein (GFP) from 73 ± 4.0% to 126 ± 3.0% and tdTomato from 60 ± 3.3% to 117 ± 2.8%. Combined with the whole-brain imaging system, we acquired the cytoarchitectonic information and the circuit information such as individual axon and boutons which were labeled with multiple fluorescent proteins. This method shows great potential in the study of continuous fine microstructure information in large-volume tissues from different species, which can facilitate the neuroscience research and help the understanding of the structure-function relationship in complex bio-tissues.

Keywords: Biomedical Materials; Techniques in Neuroscience; Tissue Engineering.