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Gold Nanoparticle Uptake in Whole Cells in Liquid Examined by Environmental Scanning Electron Microscopy

Published online by Cambridge University Press:  21 January 2014

Diana B. Peckys  and
Niels de Jonge
Diana B. Peckys*
Affiliation:
Innovative Electron Microscopy Group, INM-Leibniz Institute for New Materials, 66123 Saarbrücken, Germany
Niels de Jonge
Affiliation:
Innovative Electron Microscopy Group, INM-Leibniz Institute for New Materials, 66123 Saarbrücken, Germany Department of Molecular Physiology and Biophysics, Vanderbilt University Medical Center, Nashville, TN 37232, USA
*
*Corresponding author. E-mail: [email protected]
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Abstract

The size of gold nanoparticles (AuNPs) can influence various aspects of their cellular uptake. Light microscopy is not capable of resolving most AuNPs, while electron microscopy (EM) is not practically capable of acquiring the necessary statistical data from many cells and the results may suffer from various artifacts. Here, we demonstrate the use of a fast EM method for obtaining high-resolution data from a much larger population of cells than is usually feasible with conventional EM. A549 (human lung carcinoma) cells were subjected to uptake protocols with 10, 15, or 30 nm diameter AuNPs with adsorbed serum proteins. After 20 min, 24 h, or 45 h, the cells were fixed and imaged in whole in a thin layer of liquid water with environmental scanning electron microscopy equipped with a scanning transmission electron microscopy detector. The fast preparation and imaging of 145 whole cells in liquid allowed collection of nanoscale data within an exceptionally small amount of time of ~80 h. Analysis of 1,041 AuNP-filled vesicles showed that the long-term AuNP storing lysosomes increased their average size by 80 nm when AuNPs with 30 nm diameter were uptaken, compared to lysosomes of cells incubated with AuNPs of 10 and 15 nm diameter.

Keywords

whole cellsenvironmental scanning electron microscopyscanning transmission electron microscopygold nanoparticleslysosomesnanoparticle uptakenanoparticle size
Type
Biological Applications
Information
Microscopy and Microanalysis , Volume 20 , Issue 1 , February 2014 , pp. 189 - 197
Copyright
Copyright © Microscopy Society of America 2014 

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