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A Correlative Analysis of Gold Nanoparticles Internalized by A549 Cells
Author(s) -
Böse Katharina,
Koch Marcus,
Cavelius Christian,
Kiemer Alexandra K.,
Kraegeloh Annette
Publication year - 2014
Publication title -
particle and particle systems characterization
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 56
eISSN - 1521-4117
pISSN - 0934-0866
DOI - 10.1002/ppsc.201300308
Subject(s) - colloidal gold , fluorescence , nanoparticle , microscopy , transmission electron microscopy , confocal , fluorescence microscope , particle (ecology) , materials science , nanotechnology , confocal microscopy , electron microscope , scanning electron microscope , chemistry , biophysics , analytical chemistry (journal) , optics , chromatography , biology , physics , ecology , composite material
Fluorescently labeled nanoparticles are widely used to investigate nanoparticle cell interactions by fluorescence microscopy. Owing to limited lateral and axial resolution, nanostructures (<100 nm) cannot be resolved by conventional light microscopy techniques. Especially after uptake into cells, a common fate of the fluorescence label and the particle core cannot be taken for granted. In this study, a correlative approach is presented to image fluorescently labeled gold nanoparticles inside whole cells by correlative light and electron microscopy (CLEM). This approach allows for detection of the fluorescently labeled particle shell as well as for the gold core in one sample. In this setup, A549 cells are exposed to 8 nm Atto 647N‐labeled gold nanoparticles (3.3 × 10 9 particles mL −1 , 0.02 μg Au mL −1 ) for 5 h and are subsequently imaged by confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). Eight fluorescence signals located at different intracellular positions are further analyzed by TEM. Five of the eight fluorescence spots are correlated with isolated or agglomerated gold nanoparticles. Three fluorescence signals could not be related to the presence of gold, indicating a loss of the particle shell.