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Application of Cryogenic Transmission Electron Microscopy for Evaluation of Vaccine Delivery Carriers

  • Hui Qian
  • Yimei Jia
  • Michael J. McCluskieEmail author
Protocol
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Part of the Methods in Molecular Biology book series (MIMB, volume 2183)

Abstract

Cryogenic transmission electron microscopy (Cryo-TEM) enables visualizing the physicochemical structure of nanocarriers in solution. Here, we demonstrate the typical applications of Cryo-TEM in characterizing archaeosome-based vesicles as antigen carriers, including the morphology and size of vaccine carriers. Cryo-TEM tomography, incorporated with immunogold labeling for identifying and localizing the antigens, reveals the antigen distribution within archaeosomes in three dimensions (3D).

Key words

Cryo-TEM Plunge freezing Electron Tomography Immunogold labeling Vaccine delivery carriers 

Notes

Acknowledgments

This work was supported by the Nanotechnology Research Center and the Human Health Therapeutics Research Center at the National Research Council of Canada.

References

  1. 1.
    Dubochet J et al (1988) Cryo-electron microscopy of vitrified specimens. Q Rev Biophys 21(2):129–228CrossRefGoogle Scholar
  2. 2.
    Bartesaghi A et al (2018) Atomic resolution cryo-EM structure of β-galactosidase. Structure 26:848CrossRefGoogle Scholar
  3. 3.
    Merk A et al (2016) Breaking cryo-EM resolution barriers to facilitate drug discovery. Cell 165:1698–1707CrossRefGoogle Scholar
  4. 4.
    Kuntsche J et al (2011) Cryogenic transmission electron microscopy (cryo-TEM) for studying the morphology of colloidal drug delivery systems. Int J Pharm 417:120–137CrossRefGoogle Scholar
  5. 5.
    Venkataraman S et al (2011) The effects of polymeric nanostructure shape on drug delivery. Adv Drug Deliv Rev 63:1228–1246CrossRefGoogle Scholar
  6. 6.
    Paul AL et al (2012) Immunoelectron microscopy: a reliable tool for the analysis of cellular processes. In: Applications of immunocytochemistry. InTech, Rijeka, Croatia, pp 65–96Google Scholar
  7. 7.
    Jia Y et al (2019) A comparison of the immune responses induced by antigens in three different archaeosome-based vaccine formulations. Int J Pharm 561:187–196CrossRefGoogle Scholar
  8. 8.
    Whitfield DM et al (2016) Sulfated-glycolipids as adjuvants for vaccines. National Research Council of Canada. Patent No. WO/2016/004512Google Scholar
  9. 9.
    Aebi U et al (1987) A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J Electron Microsc Tech 7:29–33CrossRefGoogle Scholar
  10. 10.
    Egerton RF (2013) Control of radiation damage in the TEM. Ultramicroscopy 127:100–108CrossRefGoogle Scholar
  11. 11.
    Glaeser RM (1971) Limitations to significant information in biological electron microscopy as a result of radiation damage. J Ultrastruct Res 36:466–482CrossRefGoogle Scholar
  12. 12.
    Lawrence MC (1992) Least-squares methods of alignment using markers. In: Frank J (ed) Electron tomography. Plenum, New York, pp 197–204CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2021

Authors and Affiliations

  1. 1.Nanotechnology Research CenterNational Research Council CanadaEdmontonCanada
  2. 2.Human Health TherapeuticsNational Research Council CanadaOttawaCanada

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