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Abstract Osmium-based staining methods have been widely employed to enhance tissue contrast and electrical conductivity, which are crucial for the imaging throughput of the newly emerging volume electron microscopy (VEM). However, robust preparation of densely and uniformly stained tissue blocks remains a major challenge for large-scale, VEM-based ultrastructure analysis, such as connectomics. In this webinar, I will review the history of protocol development for large tissues and discuss the staining mechanism based on collective evidence. Lastly, I will share our recent experience in preparing a variety of rodent organs for X-ray microscopy and VEM. Biography Dr Yunfeng Hua studied Chemistry in Shanghai (Tongji University, B.Sc) and Munich (LMU, M.Sc). In 2012, he received a PhD degree in Neurobiology at the Max-Planck Institute of Biophysical Chemistry under the supervision of Prof. Erwin Neher. Thereafter, he joined the team of Prof. Moritz Helmstaedter (Max-Planck Institute for Brain Research) as a post-doctoral researcher to work on connectomics. In 2018, he established his own group at the Shanghai Institute of Precision Medicine with a research focus on auditory neural circuits. He has authored more than 20 scientific papers in peer-reviewed journals, including Nature Neuroscience, Neuron, Nature Communications, PNAS, and Cell Reports. Volume Imaging Australia This webinar is presented by Volume Imaging Australia, a special interest group of the Australian Microscopy and Microanalysis Society (AMMS), and Microscopy Australia.