Immunoelectron Microscopy for Visualization of Nanoparticles

  • Sarah R. Anderson
  • David Parmiter
  • Ulrich Baxa
  • Kunio NagashimaEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1682)


Immunoelectron microscopy (IEM) on a solid phase such as a carbon film is a fast and powerful way to detect and visualize surface antigens on nanoparticles by using a transmission electron microscope (TEM). Nanoparticles, in particular ones for medical applications, are often modified on the surface with soft materials to make them more soluble, less toxic, or targetable to cancerous tumors. Imaging the soft material on the surface of solid nanoparticles by electron microscopy is often a challenge. IEM can overcome this issue in cases where antibodies to any of the surface material are available, which is often the case for proteins, but also for commonly used materials such as polyethylene glycol (PEG). This effective procedure has been used traditionally for viruses and macromolecules, but it can be directly applied to nanoparticles.

Key words

Electron microscope Negative stain Immune electron microscopy Indirect solid phase immunolabeling Nanomaterial 



This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.


  1. 1.
    McNeil SE (2005) Nanotechnology for the biologist. J Leukoc Biol 78(3):585–594. doi: 10.1189/jlb.0205074 CrossRefPubMedGoogle Scholar
  2. 2.
    Hu M, Qian L, Brinas RP, Lymar ES, Hainfeld JF (2007) Assembly of nanoparticle-protein binding complexes: from monomers to ordered arrays. Angew Chem Int Ed Engl 46(27):5111–5114. doi: 10.1002/anie.200701180 CrossRefPubMedGoogle Scholar
  3. 3.
    Lee YK, Choi EJ, Webster TJ, Kim SH, Khang D (2015) Effect of the protein corona on nanoparticles for modulating cytotoxicity and immunotoxicity. Int J Nanomedicine 10:97–113. doi: 10.2147/ijn.s72998 PubMedGoogle Scholar
  4. 4.
    Anderson SR, Smith MC, Clogston JD, Patri AK, McNeil SE, Baxa U (2015) Imaging of polyethylene glycol layers on nanoparticles. Microsc Microanal 21(Suppl 3):231–232. doi: 10.1017/S1431927615001956 CrossRefGoogle Scholar
  5. 5.
    Lee Z, Jeon KJ, Dato A, Erni R, Richardson TJ, Frenklach M, Radmilovic V (2009) Direct imaging of soft-hard interfaces enabled by graphene. Nano Lett 9(9):3365–3369. doi: 10.1021/nl901664k CrossRefPubMedGoogle Scholar
  6. 6.
    Doane FW, Anderson N (1987) Electron microscopy in diagnostic virology: a practical guide and atlas. Cambridge University Press, LondonGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  • Sarah R. Anderson
    • 2
  • David Parmiter
    • 1
  • Ulrich Baxa
    • 1
  • Kunio Nagashima
    • 1
    Email author
  1. 1.Cancer Research Technology Program, Electron Microscopy LaboratoryLeidos Biomedical Research, Inc., Frederick National Laboratory for Cancer ResearchFrederickUSA
  2. 2.Microscopic Imaging Lab, Global Pathology, Drug Safety Research and DevelopmentPfizer, Inc.GrotonUSA

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