Near‐Freezing‐Temperature Golgi Neuronal Staining for X‐ray Imaging of Human Brain
Source abstract
Abstract
Abstract Achieving detailed neuronal structural information in large‐volume brain tissue has been a longstanding challenge in human brain imaging. A key obstacle arises from the trade‐off between staining efficiency and tissue autolysis. Traditional Golgi staining, typically conducted at room temperature or 37 °C to optimize staining efficiency, leads to rapid autolysis of brain tissue, resulting in the loss of fine structural details. Here, a near‐freezing temperature (NFT) staining strategy in post‐mortem frozen (PMF) human brain samples are presented, using a mercury chloride‐based method under ice‐water bath conditions. In contrast to the 37 °C Golgi staining, this NFT‐based method significantly reduces tissue autolysis, preserving fine neuronal structures. Notably, neuronal counts in the same field of view increased by 5.5‐fold, and dendritic spine density increases by 22‐fold. Using this approach, uniform staining of millimeter‐thick is achieved, centimeter‐scale human brain slices and integrated it with synchrotron‐based X‐ray microscopy to perform micrometer resolution 3D reconstructions of the cerebellum and frontal lobe. This novel technique offers a powerful tool for the fine‐structural imaging of large‐volume brain tissue, providing new insights into the intricate organization of neural networks.
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Research context
OpenAlex topics: Advanced Electron Microscopy Techniques and Applications · Advanced Fluorescence Microscopy Techniques · Molecular Biology Techniques and Applications
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
Affiliations
CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics Chinese Academy of Sciences Shanghai 201800 China
University of Chinese Academy of Sciences Beijing 100049 China
Institute of Materiobiology, College of Sciences Shanghai University Shanghai 200444 China
Xiangfu Laboratory Jiaxing 314102 China
National Human Brain Bank for Development and Function, School of Basic Medicine Peking Union Medical College Institute of Basic Medical Sciences Chinese Academy of Medical Sciences Beijing 100005 China
Shanghai Synchrotron Radiation Facility, Shanghai Advanced Research Institute Chinese Academy of Sciences Shanghai 201210 China
State Key Laboratory of Synergistic Chem‐Bio Synthesis, School of Chemistry and Chemical Engineering, New Cornerstone Science Laboratory, Frontiers Science Center for Transformative Molecules, Zhang Jiang Institute for Advanced Study and National Center for Translational Medicine Shanghai Jiao Tong University Shanghai 200240 China
Funding
National Key Research and Development Program of China · National Natural Science Foundation of China
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