Applied Microscopy

Korean Society of Microscopy (한국현미경학회)
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Structural Analysis of Exosomes Using Different Types of Electron Microscopy

  • Choi, Hyosun (BK21 Plus Program, Department of Senior Healthcare, Graduate School, Eulji University) ;
  • Mun, Ji Young (BK21 Plus Program, Department of Senior Healthcare, Graduate School, Eulji University)
  • Received : 2017.06.12
  • Accepted : 2017.07.24
  • Published : 2017.09.30
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Abstract

Negative staining has been traditionally used for exosome imaging; however, the technique is limited to surface topology only and can cause staining artifacts. Therefore, to analyze the internal structure of exosomes, we employed a method of block preparation, thin sectioning, and electron tomography. In addition, an automatic serial sectioning technique with 15-nm thickness through focused ion beam was employed to observe the three-dimensional structure of exosomes of various sizes. Cryo-transmission electron microscopy revealed the near-to-native structure of exosomes.

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References

  1. Choi H, Jung M K, and Mun J Y (2017) Current status of automatic serial sections for 3D electron microscopy. Appl. Microsc. 47, 3-7. https://doi.org/10.9729/AM.2017.47.1.3
  2. Enderle D, Spiel A, Coticchia C M, Berghoff E, Mueller R, Schlumpberger M, Sprenger-Haussels M, Shaffer J M, Lader E, Skog J, and Noerholm M (2015) Characterization of RNA from exosomes and other extracellular vesicles isolated by a novel spin column-based method. PLoS One 10, e0136133. https://doi.org/10.1371/journal.pone.0136133
  3. Gyorgy B, Szabo T G, Pasztoi M, Pal Z, Misjak P, Aradi B, Laszlo V, Pallinger E, Pap E, Kittel A, Nagy G, Falus A, and Buzas E I (2011) Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles. Cell. Mol. Life Sci. 68, 2667-2688. https://doi.org/10.1007/s00018-011-0689-3
  4. Jimenez N, Humbel B M, van Donselaar E, Verkleij A J, and Burger K N (2006) Aclar discs: a versatile substrate for routine high-pressure freezing of mammalian cell monolayers. J. Microsc. 221, 216-223. https://doi.org/10.1111/j.1365-2818.2006.01558.x
  5. Kim S B, Kim H R, Park M C, Cho S, Goughnour P C, Han D, Yoon I, Kim Y, Kang T, Song E, Kim P, Choi H, Mun J Y, Song C, Lee S, Jung H S, and Kim S (2017) Caspase-8 controls the secretion of inflammatory lysyltRNA synthetase in exosomes from cancer cells. J. Cell Biol. 216, 2201-2216. https://doi.org/10.1083/jcb.201605118
  6. Liu X and Wang H W (2011) Single particle electron microscopy reconstruction of the exosome complex using the random conical tilt method. J. Vis. Exp. doi: 10.3791/2574.
  7. Montaner-Tarbes S, Borras F E, Montoya M, Fraile L, and Del Portillo H A (2016) Serum-derived exosomes from non-viremic animals previously exposed to the porcine respiratory and reproductive virus contain antigenic viral proteins. Vet. Res. 47, 59. https://doi.org/10.1186/s13567-016-0345-x
  8. Raposo G and Stoorvogel W (2013) Extracellular vesicles: exosomes, microvesicles, and friends. J. Cell Biol. 200, 373-383. https://doi.org/10.1083/jcb.201211138
  9. Saari H, Lazaro-Ibanez E, Viitala T, Vuorimaa-Laukkanen E, Siljander P, and Yliperttula M (2015) Microvesicle- and exosome-mediated drug delivery enhances the cytotoxicity of Paclitaxel in autologous prostate cancer cells. J. Control. Release 220, 727-737. https://doi.org/10.1016/j.jconrel.2015.09.031
  10. Sharma S, Rasool H I, Palanisamy V, Mathisen C, Schmidt M, Wong D T, and Gimzewski J K (2010) Structural-mechanical characterization of nanoparticle exosomes in human saliva, using correlative AFM, FESEM, and force spectroscopy. ACS Nano 4, 1921-1926. https://doi.org/10.1021/nn901824n
  11. Thery C (2011) Exosomes: secreted vesicles and intercellular communications. F1000 Biol. Rep. 3, 15.
  12. Thery C, Ostrowski M, and Segura E (2009) Membrane vesicles as conveyors of immune responses. Nat. Rev. Immunol. 9, 581-593. https://doi.org/10.1038/nri2567
  13. Titze B and Genoud C (2016) Volume scanning electron microscopy for imaging biological ultrastructure. Biol. Cell 108, 307-323. https://doi.org/10.1111/boc.201600024
  14. Villarroya-Beltri C, Baixauli F, Gutierrez-Vazquez C, Sanchez-Madrid F, and Mittelbrunn M (2014) Sorting it out: regulation of exosome loading. Semin. Cancer Biol. 28, 3-13. https://doi.org/10.1016/j.semcancer.2014.04.009
  15. Vlassov A V, Magdaleno S, Setterquist R, and Conrad R (2012) Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim. Biophys. Acta 1820, 940-948. https://doi.org/10.1016/j.bbagen.2012.03.017
  16. Zaborowski M P, Balaj L, Breakefield X O, and Lai C P (2015) Extracellular vesicles: composition, biological relevance, and methods of study. Bioscience 65, 783-797. https://doi.org/10.1093/biosci/biv084