Observation of intracellular protein localization area in a single neuron using gold nanoparticles with a scanning electron microscope

•Protein localization areas in a neuron were observed with SEM.•Proteins were immunologically stained with gold nanoparticles and visualized.•Protein localization areas in SEM images agree well with those found in optical images.•Protein localization areas would not be modified after sample preparat...

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Veröffentlicht in:Micron (Oxford, England : 1993) England : 1993), 2019-11, Vol.126, p.102740-102740, Article 102740
Hauptverfasser: Goto, Toichiro, Kasai, Nahoko, Filip, Roxana, Sumitomo, Koji, Nakashima, Hiroshi
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Sprache:eng
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Zusammenfassung:•Protein localization areas in a neuron were observed with SEM.•Proteins were immunologically stained with gold nanoparticles and visualized.•Protein localization areas in SEM images agree well with those found in optical images.•Protein localization areas would not be modified after sample preparation. The localization areas of intracellular proteins in rat cortical neurons were visualized using a scanning electron microscope (SEM) coupled with a focused ion beam (FIB) system. To obtain a clear contrast in the SEM images, gold nanoparticles (GNPs) were bound to specific intracellular proteins by antigen-antibody reactions. By obtaining a cross section of the desired location of the neurons by FIB milling under the SEM imaging condition, it was possible to observe the proteins inside the cells as clear bright spots. When a neuron was stained with anti-tau and anti-histone H1 antibodies, the bright spots were localized in the cross section of the axon and the nucleus, respectively. It was confirmed that targeted proteins in a single neuron on a substrate could be successfully identified. The development of FIB/SEM observation with immunological GNP staining will offer important information for the stable growth of neurons on various substrate structures, since the elongation and turning of axons on the substrates are activated by the redistribution of intracellular proteins.
ISSN:0968-4328
1878-4291
DOI:10.1016/j.micron.2019.102740