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- W2801065963 abstract "Immunogold labeling of permeabilized whole-mount cells or thin-sectioned material is widely used for the subcellular localization of biomolecules at the high spatial resolution of electron microscopy (EM). Those approaches are well compatible with either 3-dimensional (3D) reconstruction of organelle morphology and antigen distribution or with rapid cryofixation-but not easily with both at once. We describe here a specimen preparation and labeling protocol for animal cell cultures, which represents a novel blend of specifically adapted versions of established techniques. It combines the virtues of reliably preserved organelle ultrastructure, as trapped by rapid freezing within milliseconds followed by freeze-substitution and specimen rehydration, with the advantages of robust labeling of intracellular constituents in 3D through means of pre-embedding NANOGOLD-silver immunocytochemistry. So obtained thin and semi-thick epoxy resin sections are suitable for transmission EM imaging, as well as tomographic reconstruction and modeling of labeling patterns in the 3D cellular context." @default.
- W2801065963 created "2018-05-17" @default.
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- W2801065963 date "2018-05-21" @default.
- W2801065963 modified "2023-10-01" @default.
- W2801065963 title "Combining high-pressure freezing with pre-embedding immunogold electron microscopy and tomography" @default.
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- W2801065963 doi "https://doi.org/10.1111/tra.12575" @default.
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