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High Initial Sputter Rate Found for Vaccinia Virions Using Isotopic Labeling, NanoSIMS, and AFM

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...

Authors:   Sean D. Gates; Richard C. Condit; Nissin Moussatche; Benjamin J. Stewart; Alexander J. Malkin; Peter K. Weber
Journal:   Analytical Chemistry
Year:   2018
DOI:   10.1021/acs.analchem.7b02786
Publication date:   19-Jan-2018
Facts, background information, dossiers
  • virions
  • DNA
  • particles
  • imaging
  • glutaraldehyde
More about American Chemical Society Publications
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