Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-10T13:51:40.402Z Has data issue: false hasContentIssue false
  • English
  • Français

A Scanning Transmission Electron Microscopy Approach to Analyzing Large Volumes of Tissue to Detect Nanoparticles

Published online by Cambridge University Press:  27 June 2013

Paul J. Kempen ,
Avnesh S. Thakor ,
Cristina Zavaleta ,
Sanjiv S. Gambhir  and
Robert Sinclair
Paul J. Kempen*
Affiliation:
Department of Materials Science and Engineering, Stanford University, Stanford, CA 94305-4034, USA
Avnesh S. Thakor
Affiliation:
Molecular Imaging Program at Stanford, Department of Radiology, Stanford University, Stanford, CA 94305-5427, USA Department of Radiology, University of Cambridge, Cambridge CB2 2QQ, UK
Cristina Zavaleta
Affiliation:
Molecular Imaging Program at Stanford, Department of Radiology, Stanford University, Stanford, CA 94305-5427, USA
Sanjiv S. Gambhir
Affiliation:
Department of Materials Science and Engineering, Stanford University, Stanford, CA 94305-4034, USA Molecular Imaging Program at Stanford, Department of Radiology, Stanford University, Stanford, CA 94305-5427, USA Department of Bio-engineering and Bio-X Program, Stanford University, Stanford, CA 94305-5427, USA
Robert Sinclair
Affiliation:
Department of Materials Science and Engineering, Stanford University, Stanford, CA 94305-4034, USA
*
*Corresponding author. E-mail: pkempen@stanford.edu
Get access

Abstract

The use of nanoparticles for the diagnosis and treatment of cancer requires the complete characterization of their toxicity, including accurately locating them within biological tissues. Owing to their size, traditional light microscopy techniques are unable to resolve them. Transmission electron microscopy provides the necessary spatial resolution to image individual nanoparticles in tissue, but is severely limited by the very small analysis volume, usually on the order of tens of cubic microns. In this work, we developed a scanning transmission electron microscopy (STEM) approach to analyze large volumes of tissue for the presence of polyethylene glycol-coated Raman-active-silica-gold-nanoparticles (PEG-R-Si-Au-NPs). This approach utilizes the simultaneous bright and dark field imaging capabilities of STEM along with careful control of the image contrast settings to readily identify PEG-R-Si-Au-NPs in mouse liver tissue without the need for additional time-consuming analytical characterization. We utilized this technique to analyze 243,000 μm3 of mouse liver tissue for the presence of PEG-R-Si-Au-NPs. Nanoparticles injected into the mice intravenously via the tail vein accumulated in the liver, whereas those injected intrarectally did not, indicating that they remain in the colon and do not pass through the colon wall into the systemic circulation.

Keywords

STEMnanotechnologycancertissue analysislarge volume analysis
Type
Techniques and Software Development
Information
Microscopy and Microanalysis , Volume 19 , Issue 5 , October 2013 , pp. 1290 - 1297
Copyright
Copyright © Microscopy Society of America 2013 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Chang, E., Thekkek, N., Yu, W.W., Colvin, V.L. & Drezek, R. (2006). Evaluation of quantum dot cytotoxicity based on intracellular uptake. Small 2(12), 1412–1417.CrossRefGoogle ScholarPubMed
Devaraj, N.K., Keliher, E.J., Thurber, G.M., Nahrendorf, M. & Weissleder, R. (2009). 18F labeled nanoparticles for in vivo PET-CT imaging. Bioconjugate Chem 20(2), 397–401.Google Scholar
Farrell, D., Alper, J., Ptak, K., Panaro, N.J., Grodzinski, P. & Barker, A.D. (2010). Recent advances from the national cancer institute alliance for nanotechnology in cancer. ACS Nano 4(2), 589–594.Google Scholar
Ferrari, M. (2005). Cancer nanotechnology: Opportunities and challenges. Nat Rev Cancer 5(3), 161–171.Google Scholar
Inderbitzin, D., Gass, M., Beldi, G., Ayouni, E., Nordin, A., Sidler, D., Gloor, B., Candinas, D. & Stoupis, C. (2004). Magnetic resonance imaging provides accurate and precise volume determination of the regenerating mouse liver. J Gastrointest Surg 8(7), 806–811.Google Scholar
Keren, S., Zavaleta, C., Cheng, Z., de la Zerda, A., Gheysens, O. & Gambhir, S.S. (2008). Noninvasive molecular imaging of small living subjects using Raman spectroscopy. Proc Natl Acad Sci U S A 105(15), 5844–5849.Google Scholar
Kircher, M.F., de la Zerda, A., Jokerst, J.V., Zavaleta, C.L., Kempen, P.J., Mittra, E., Pitter, K., Huang, R., Campos, C., Habte, F., Sinclair, R., Brennan, C.W., Mellinghoff, I.K., Holland, E.C. & Gambhir, S.S. (2012). A brain tumor molecular imaging strategy using a new triple-modality MRI-photoacoustic-Raman nanoparticle. Nat Med 18(5), 829–834.Google Scholar
Kneipp, J., Kneipp, H., Rajadurai, A., Redmond, R.W. & Kneipp, K. (2009). Optical probing and imaging of live cells using SERS labels. J Raman Spectrosc 40(1), 1–5.Google Scholar
Koh, A.L., Shachaf, C.M., Elchuri, S., Nolan, G.P. & Sinclair, R. (2008). Electron microscopy localization and characterization of functionalized composite organic-inorganic SERS nanoparticles on leukemia cells. Ultramicroscopy 109(1), 111–121.Google Scholar
Liang, G., Ronald, J., Chen, Y., Ye, D., Pandit, P., Ma, M.L., Rutt, B. & Rao, J. (2011). Controlled self-assembling of gadolinium nanoparticles as smart molecular magnetic resonance imaging contrast agents. Angew Chemie, Int Ed 50(28), 6283–6286.Google Scholar
Liu, Z., Tabakman, S., Sherlock, S., Li, X., Chen, Z., Jiang, K., Fan, S. & Dai, H. (2010). Multiplexed five-color molecular imaging of cancer cells and tumor tissues with carbon nanotube Raman tags in the near-infrared. Nano Res 3(3), 222–233.Google Scholar
Loo, C., Lowery, A., Halas, N., West, J. & Drezek, R. (2005). Immunotargeted nanoshells for integrated cancer imaging and therapy. Nano Lett 5(4), 709–711.Google Scholar
Mulvaney, S.P., Musick, M.D., Keating, C.D. & Natan, M.J. (2003). Glass-coated, analyte-tagged nanoparticles: A new tagging system based on detection with surface-enhanced raman scattering. Langmuir 19 (11), 4784–4790.Google Scholar
Panessa-Warren, B.J., Warren, J.B., Wong, S.S. & Misewich, J.A. (2006). Biological cellular response to carbon nanoparticle toxicity. J Phys: Condens Matter 18(33), S2185. Google Scholar
Ptak, K., Farrell, D., Panaro, N.J., Grodzinski, P. & Barker, A.D. (2010). The NCI alliance for nanotechnology in cancer: Achievement and path forward. Wiley Interdiscip rev: Nanomed Nanobiotechnol 2(5), 450–460.Google Scholar
Thakor, A.S., Jokerst, J., Zavaleta, C., Massoud, T.F., Gambhir, S.S. (2011a). Gold nanoparticles: A revival in precious metal administration to patients. Nano Lett 11(10), 4029–4036.CrossRefGoogle ScholarPubMed
Thakor, A.S., Luong, R., Paulmurugan, R., Lin, F.I., Kempen, P., Zavaleta, C., Chu, P., Massoud, T.F., Sinclair, R. & Gambhir, S.S. (2011b). The fate and toxicity of Raman-active silica-gold nanoparticles in mice. Sci Transl Med 3(79), 79ra33. Google Scholar
Thakor, A.S., Paulmurugan, R., Kempen, P., Zavaleta, C., Sinclair, R., Massoud, T.F. & Gambhir, S.S. (2011c). Oxidative stress mediates the effects of Raman-active gold nanoparticles in human cells. Small 7(1), 126–136.Google Scholar
von Maltzahn, G., Centrone, A., Park, J.H., Ramanathan, R., Sailor, M.J., Hatton, T.A. & Bhatia, S.N. (2009). SERS-coded gold nanorods as a multifunctional platform for densely multiplexed near-infrared imaging and photothermal heating. Adv Mater 21(31), 3175–3180.CrossRefGoogle ScholarPubMed
Weibel, E.R. (1979). Stereological Methods, pp. 114–115. New York: Academic Press.Google Scholar
Williams, D.B. & Carter, C.B. (2009). Transmission Electron Microscopy: A Textbook for Materials Science, vol. 9, p. 379. New York: Springer.CrossRefGoogle Scholar
Zavaleta, C.L., Smith, B.R., Walton, I., Doering, W., Davis, G., Shojaei, B., Natan, M.J. & Gambhir, S.S. (2009). Multiplexed imaging of surface enhanced Raman scattering nanotags in living mice using noninvasive Raman spectroscopy. Proc Natl Acad Sci USA 106(32), 13511–13516.CrossRefGoogle ScholarPubMed