Correlative light and electron microscopy imaging of proteinaceous deposits in cell cultures and brain tissues.

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Tác giả: Dennis W Dickson, Peizhou Jiang

Ngôn ngữ: eng

Ký hiệu phân loại: 612.826 Diencephalon and brain stem

Thông tin xuất bản: England : Acta neuropathologica communications , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 693358

Identifying protein deposits and associated components is crucial for understanding the pathogenesis of neurodegenerative disorders with intracellular or extracellular deposits. Correlative light and electron microscopy (CLEM) has emerged as a powerful tool to accurately study tissue and cellular pathology by examination of the same target at both microstructural and ultrastructural levels. However, the technical challenges with CLEM have limited its application to neuropathology. Here, we developed a simplified efficient CLEM method and applied it to a cell model that produces a high proportion of α-synuclein (αS) inclusions with immunopositivity to phosphorylated αS and the synaptic vesicle marker SV2A and synaptophysin. This approach incorporates modifications in sample processing and innovative fiducial marking techniques, which enhance antigen preservation and improve target registration, respectively. These advancements achieve an optimal balance in sensitivity, accuracy, efficiency, and cost-effectiveness compared to current CLEM methods employing different strategies. Using this method, we identified and analyzed αS inclusions in cell cultures, as well as various pathological protein deposits in postmortem brain tissues from individuals with a range of neurodegenerative disorders. Our findings replicate recently reported new features of αS pathology and also reveal unrecognized a variety forms of small αS inclusions in human brain, which provide valuable insights into mechanisms underlying Lewy-related pathology. Application of this enhanced CLEM method is a powerful tool in research on neurodegenerative disorders, including αS-opathies.
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