Open Access

Imaging of immunogold labeling in cells and tissues by helium ion microscopy

  • Authors:
    • Chikara Sato
    • Mari Sato
    • Shinichi Ogawa
  • View Affiliations

  • Published online on: March 30, 2018     https://doi.org/10.3892/ijmm.2018.3604
  • Pages: 309-321
  • Copyright: © Sato et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Helium ion microscopy (HIM) scans samples with a fine ion beam exploiting the very short de Broglie wavelength of helium ions. Because the radiation induces only a small sample region to emit secondary electrons (SEs), very high resolution is expected. In order to explore the applications of SE-HIM in biology, COS7 kidney fibroblast cells and C2C12 myoblast cells cultured on a silicon (Si) nitride (SiN)/Si bilayer were dried and directly observed in high vacuum, without coating or staining. High contrast, high depth-of-field images were obtained revealing the nucleus, endoplasmic reticulum, cytoskeleton and putative mitochondria above a bright background from the support. Gold-tagged antibodies were employed to aid organelle identification. Signals from the gold tags were most clearly distinguishable by secondary electron (SE)-HIM when cells were grown on thin SiN film, and the minimum gap measured between gold particles showed the resolution to be 2 nm. Wheat germ agglutinin-gold labeling revealed clusters of gold particles ~50-200 nm in diameter on COS7 cells, which might represent assemblies of glycosylated proteins, suggesting the formation of membrane raft structures that include membrane proteins. SE-HIM also delivered high contrast images of unstained, uncoated, thin sections of Epon‑embedded mouse kidney tissues mounted on a SiN/Si bilayer, revealing the details of sub-tissues and cell organelles. A charge-coupled mechanism explaining the observed SE-HIM contrast is proposed. Ionoluminescence-HIM was also performed targeting zinc oxide particles on cells. In conclusion, the high depth-of-field, high-resolution imaging achieved using HIM may have applications in various fields, including soft materials.
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July-2018
Volume 42 Issue 1

Print ISSN: 1107-3756
Online ISSN:1791-244X

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Spandidos Publications style
Sato C, Sato M and Ogawa S: Imaging of immunogold labeling in cells and tissues by helium ion microscopy. Int J Mol Med 42: 309-321, 2018
APA
Sato, C., Sato, M., & Ogawa, S. (2018). Imaging of immunogold labeling in cells and tissues by helium ion microscopy. International Journal of Molecular Medicine, 42, 309-321. https://doi.org/10.3892/ijmm.2018.3604
MLA
Sato, C., Sato, M., Ogawa, S."Imaging of immunogold labeling in cells and tissues by helium ion microscopy". International Journal of Molecular Medicine 42.1 (2018): 309-321.
Chicago
Sato, C., Sato, M., Ogawa, S."Imaging of immunogold labeling in cells and tissues by helium ion microscopy". International Journal of Molecular Medicine 42, no. 1 (2018): 309-321. https://doi.org/10.3892/ijmm.2018.3604