biostudies-literatureZhou FSTI2030-Major ProjectNational Key R&D Program of China2022 Shanghai "Science and Technology Innovation Action Plan" Fundamental Research ProjectNational Natural Science Foundation of ChinaNational Key Research and Development Program of Chinae04468https://www.ebi.ac.uk/biostudies/studies/S-EPMC12376569biostudies-literatureUnknown12(30)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 Advanced science (Weinheim, Baden-Wurttemberg, Germany)Near-Freezing-Temperature Golgi Neuronal Staining for X-ray Imaging of Human Brain.PMC123765692021ZD02011002022YFA160360022325406T2188102223939342199113422JC1401203Li JZhang JZhu YLi QYan XWang LMa CHu JTang QZhang YZhou FCai XFan CfalseNear-Freezing-Temperature Golgi Neuronal Staining for X-ray Imaging of Human Brain.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 2025-01-01T00:00:00Z2025 Aug2026-05-09T19:08:06.951Z2026-04-08T01:09:49.655ZS-EPMC123765694043405210.1002/advs.202504468