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ACS Publications. Most Trusted. Most Cited. Most Read
High Initial Sputter Rate Found for Vaccinia Virions Using Isotopic Labeling, NanoSIMS, and AFM

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Download Hi-Res ImageDownload to MS-PowerPointCite This:Anal. Chem. 2018, 90, 3, 1613-1620
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    High Initial Sputter Rate Found for Vaccinia Virions Using Isotopic Labeling, NanoSIMS, and AFM
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    † †Nuclear and Chemical Sciences Division and §Biosciences and Biotechnology Division, Lawrence Livermore National Laboratory, Livermore, California 94551, United States
    ‡ Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, Florida 32610, United States
    *E-mail for A.J.M.: [email protected]. Tel.: 925-423-7817.
    *E-mail for P.K.W.: [email protected]. Tel.: 925-422-3018.
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    Analytical Chemistry

    Cite this: Anal. Chem. 2018, 90, 3, 1613–1620
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    https://doi.org/10.1021/acs.analchem.7b02786
    Published January 3, 2018
    Copyright © 2018 American Chemical Society
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    Abstract

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    High-lateral-resolution secondary ion mass spectrometry (SIMS) has the potential to provide functional and depth resolved information from small biological structures, such as viral particles (virions) and phage, but sputter rate and sensitivity are not characterized at shallow depths relevant to these structures. Here we combine stable isotope labeling of the DNA of vaccinia virions with correlated SIMS imaging depth profiling and atomic force microscopy (AFM) to develop a nonlinear, nonequilibrium sputter rate model for the virions and validate the model on the basis of reconstructing the location of the DNA within individual virions. Our experiments with a Cs+ beam show an unexpectedly high initial sputter rate (∼100 um2·nm·pA–1·s–1) with a rapid decline to an asymptotic rate of 0.7 um2·nm·pA–1·s–1 at an approximate depth of 70 nm. Correlated experiments were also conducted with glutaraldehyde-fixed virions, as well as O– and Ga+ beams, yielding similar results. Based on our Cs+ sputter rate model, the labeled DNA in the virion was between 50 and 90 nm depth in the virion core, consistent with expectations, supporting our conclusions. Virion densification was found to be a secondary effect. Accurate isotopic ratios were obtained from the initiation of sputtering, suggesting that isotopic tracers could be successfully used for smaller virions and phage.

    Copyright © 2018 American Chemical Society

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    Supporting Information

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    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.analchem.7b02786.

    • Information on chemical fixation, molecular fractionation of virions, correlated AFM-SIMS analysis, orientation of dispersed virions, sputtering of iridium coated samples, heterogeneity of isotopic labeling, unlabeled virion depth profiles, gallium and oxygen ion sputtering, Cs+ sputtering of glutaraldehyde-fixed virions, rejection of vacuum-induced densification, and ion-induced densification (PDF)

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    Cited By

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    This article is cited by 8 publications.

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    2. Dong-Kyu Lee, Euiyeon Na, Seongoh Park, Jeong Hill Park, Johan Lim, Sung Won Kwon. In Vitro Tracking of Intracellular Metabolism-Derived Cancer Volatiles via Isotope Labeling. ACS Central Science 2018, 4 (8) , 1037-1044. https://doi.org/10.1021/acscentsci.8b00296
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    Analytical Chemistry

    Cite this: Anal. Chem. 2018, 90, 3, 1613–1620
    Click to copy citationCitation copied!
    https://doi.org/10.1021/acs.analchem.7b02786
    Published January 3, 2018
    Copyright © 2018 American Chemical Society
    Request reuse permissions

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