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Whole-cell analysis of low-density lipoprotein uptake by macrophages using STEM tomography

Baudoin, J.P.; Gray Jerome, W.; Kübel, C.; de Jonge, N.

Abstract:
Nanoparticles of heavy materials such as gold can be used as markers in quantitative electron microscopic studies of protein distributions in cells with nanometer spatial resolution. Studying nanoparticles within the context of cells is also relevant for nanotoxicological research. Here, we report a method to quantify the locations and the number of nanoparticles, and of clusters of nanoparticles inside whole eukaryotic cells in three dimensions using scanning transmission electron microscopy (STEM) tomography. Whole-mount fixed cellular samples were prepared, avoiding sectioning or slicing. The level of membrane staining was kept much lower than is common practice in transmission electron microscopy (TEM), such that the nanoparticles could be detected throughout the entire cellular thickness. Tilt-series were recorded with a limited tilt-range of 80u thereby preventing excessive beam broadening occurring at higher tilt angles. The 3D locations of the nanoparticles were nevertheless determined with high precision using computation. The obtained information differed from that obtained with conventional TEM tomography data since the nanoparticles were highlighted while only faint contrast was obtained on the cellular material. ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/110090503
Veröffentlicht am 06.04.2018
Originalveröffentlichung
DOI: 10.1371/journal.pone.0055022
Scopus
Zitationen: 11
Web of Science
Zitationen: 12
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Nanotechnologie (INT)
Karlsruhe Nano Micro Facility (KNMF)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2013
Sprache Englisch
Identifikator ISSN: 1932-6203
urn:nbn:de:swb:90-AAA1100905030
KITopen-ID: 110090503
HGF-Programm 43.16.02 (POF II, LK 02) KNMF laboratory for microscopy a.spect.
Erschienen in PLoS one
Verlag Public Library of Science (PLoS)
Band 8
Seiten e55022/1-8
Nachgewiesen in Scopus
Web of Science
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