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Tuijtel MW, Mulder AA, Posthuma CC, et al. Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision. Sci Rep. 2017;7(1):10442doi: 10.1038/s41598-017-10905-x.
Tuijtel, M. W., Mulder, A. A., Posthuma, C. C., van der Hoeven, B., Koster, A. J., Bárcena, M., Faas, F. G. A., & Sharp, T. H. (2017). Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision. Scientific reports, 7(1), 10442. https://doi.org/10.1038/s41598-017-10905-x
Tuijtel, Maarten W, et al. "Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision." Scientific reports vol. 7,1 (2017): 10442. doi: https://doi.org/10.1038/s41598-017-10905-x
Tuijtel MW, Mulder AA, Posthuma CC, van der Hoeven B, Koster AJ, Bárcena M, Faas FGA, Sharp TH. Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision. Sci Rep. 2017 Sep 05;7(1):10442. doi: 10.1038/s41598-017-10905-x. PMID: 28874723; PMCID: PMC5585351.
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Sci Rep. 2017 Sep 05;7(1):10442. doi: 10.1038/s41598-017-10905-x.

Inducing fluorescence of uranyl acetate as a dual-purpose contrast agent for correlative light-electron microscopy with nanometre precision.

Scientific reports

Maarten W Tuijtel, Aat A Mulder, Clara C Posthuma, Barbara van der Hoeven, Abraham J Koster, Montserrat Bárcena, Frank G A Faas, Thomas H Sharp

Affiliations

  1. Section Electron Microscopy, Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
  2. Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
  3. NeCEN, Gorlaeus Laboratories, Leiden University, 2333 CC, Leiden, The Netherlands.
  4. Section Electron Microscopy, Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands. [email protected].
  5. Section Electron Microscopy, Department of Molecular Cell Biology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands. [email protected].

PMID: 28874723 PMCID: PMC5585351 DOI: 10.1038/s41598-017-10905-x

Abstract

Correlative light-electron microscopy (CLEM) combines the high spatial resolution of transmission electron microscopy (TEM) with the capability of fluorescence light microscopy (FLM) to locate rare or transient cellular events within a large field of view. CLEM is therefore a powerful technique to study cellular processes. Aligning images derived from both imaging modalities is a prerequisite to correlate the two microscopy data sets, and poor alignment can limit interpretability of the data. Here, we describe how uranyl acetate, a commonly-used contrast agent for TEM, can be induced to fluoresce brightly at cryogenic temperatures (-195 °C) and imaged by cryoFLM using standard filter sets. This dual-purpose contrast agent can be used as a general tool for CLEM, whereby the equivalent staining allows direct correlation between fluorescence and TEM images. We demonstrate the potential of this approach by performing multi-colour CLEM of cells containing equine arteritis virus proteins tagged with either green- or red-fluorescent protein, and achieve high-precision localization of virus-induced intracellular membrane modifications. Using uranyl acetate as a dual-purpose contrast agent, we achieve an image alignment precision of ~30 nm, twice as accurate as when using fiducial beads, which will be essential for combining TEM with the evolving field of super-resolution light microscopy.

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