Journal
ULTRAMICROSCOPY
Volume 143, Issue -, Pages 93-99Publisher
ELSEVIER
DOI: 10.1016/j.ultramic.2013.09.002
Keywords
Correlative light and electron microscopy; Atmospheric electron microscopy; Scanning electron microscopy; Fluorescence microscopy; Nano particle; Quantum dots
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We investigated SEM imaging of nanoparticle biomarkers suspended below a thin membrane, with the ultimate goal of integrating functional fluorescence and structural SEM measurements of samples kept at ambient or hydrated conditions. In particular, we investigated how resolving power in liquid SEM is affected by the interaction of the electron beam with the membrane. Simulations with the Geant4-based Monte Carlo scheme developed by Kieft and Bosch (2008) 111 are compared to experimental results with suspended nanoparticles. For 20 nm and 50 nm thin membranes, we found a beam broadening of 1.5 nm and 3 nm, respectively, with an excellent agreement between simulations and experiments. 15 nm Au nanoparticles and bio-functionalizecl core-shell quantum dots can be individually resolved in denser clusters. We demonstrated the imaging of single EGF-conjugated quantum dots docked at filopodia during cellular uptake with both fluorescence microscopy and SEM simultaneously. These results open novel opportunities for correlating live fluorescence microscopy with structural electron microscopy. (C) 2013 Elsevier B.V. All rights reserved
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