title: High-throughput dual-colour precision imaging for brain-wide connectome with cytoarchitectonic landmarks at the cellular level.
abstract: The precise annotation and accurate identification of neural structures are prerequisites for studying mammalian brain function. The orientation of neurons and neural circuits is usually determined by mapping brain images to coarse axial-sampling planar reference atlases. However, individual differences at the cellular level likely lead to position errors and an inability to orient neural projections at single-cell resolution. Here, we present a high-throughput precision imaging method that can acquire a co-localized brain-wide data set of both fluorescent-labelled neurons and counterstained cell bodies at a voxel size of 0.32 × 0.32 × 2.0 μm in 3 days for a single mouse brain. We acquire mouse whole-brain imaging data sets of multiple types of neurons and projections with anatomical annotation at single-neuron resolution. The results show that the simultaneous acquisition of labelled neural structures and cytoarchitecture reference in the same brain greatly facilitates precise tracing of long-range projections and accurate locating of nuclei.
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Gong
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Dongli
authors.initials: D
authors.lastname: Xu
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yuan
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Xiangning
authors.initials: X
authors.lastname: Li
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Congdi
authors.initials: C
authors.lastname: Guo
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Peng
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Yuxin
authors.initials: Y
authors.lastname: Li
authors.affiliation: Howard Hughes Medical Institute and Department of Biology, Stanford University, 385 Serra Mall, Stanford, California 94305, USA.
authors.forename: Lindsay A
authors.initials: LA
authors.lastname: Schwarz
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Anan
authors.initials: A
authors.lastname: Li
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Bihe
authors.initials: B
authors.lastname: Hu
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Benyi
authors.initials: B
authors.lastname: Xiong
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Qingtao
authors.initials: Q
authors.lastname: Sun
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Yalun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Jiepeng
authors.initials: J
authors.lastname: Liu
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Qiuyuan
authors.initials: Q
authors.lastname: Zhong
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Tonghui
authors.initials: T
authors.lastname: Xu
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Shaoqun
authors.initials: S
authors.lastname: Zeng
authors.affiliation: Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.|Department of Biomedical Engineering, Key Laboratory for Biomedical Photonics of Ministry of Education, Huazhong University of Science and Technology, 1037 Luoyu Road, Wuhan 430074, China.
authors.forename: Qingming
authors.identifier: 0000-0002-6725-9311
authors.initials: Q
authors.lastname: Luo
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000000981:diagnostic imaging*; D003116:Color; D063132:Connectome / Q000379:methods*; D005240:Feasibility Studies; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008853:Microscopy / Q000379:methods; D023421:Models, Animal; D009434:Neural Pathways / Q000000981:diagnostic imaging*; D009474:Neurons / Q000648:ultrastructure*; D059010:Single-Cell Analysis / Q000379:methods; D014054:Tomography / Q000379:methods