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Correlative light and electron microscopy using immunolabeled resin sections.
Methods in Molecular Biology
In correlative microscopy, light microscopy provides the overview and orientation in the complex cells and tissue, whereas electron microscopy offers the detailed localization and correlation to subcellular structures. In this chapter, we offer the detailed high-quality electron microscopical preparation methods for the optimum preservation of the cellular ultrastructure. From such preparations, serial thin sections are collected and used for comparative histochemical, immunofluorescence, and immunogold staining. In light microscopy, histological stains are used to identify the orientation of the sample, and immunofluorescence labeling is used to identify the region of interest, namely, the labeled cells expressing the macromolecule under investigation. Subsequent sections, labeled with immunogold, are analyzed by electron microscopy to identify the label within the cellular architecture at high resolution.
Animals, Epoxy Resins, Fluorescent Antibody Technique, Fluorescent Dyes, Freeze Substitution, Immunohistochemistry, Intestines/metabolism, Intestines/ultrastructure, Microscopy/methods, Microscopy, Immunoelectron/methods, Plastic Embedding, Rats, Resins, Synthetic, Tissue Fixation, Trypanosoma brucei brucei/metabolism, Trypanosoma brucei brucei/ultrastructure, Zebrafish/anatomy & histology, Zebrafish/metabolism
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