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Cancer imaging using surface-enhanced resonance Raman scattering nanoparticles

Nature Protocols volume 12, pages 1400–1414 (2017)Cite this article

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Abstract

The unique spectral signatures and biologically inert compositions of surface-enhanced resonance Raman scattering (SERRS) nanoparticles make them promising contrast agents for in vivo cancer imaging. Our SERRS nanoparticles consist of a 60-nm gold nanoparticle core that is encapsulated in a 15-nm-thick silica shell wherein the resonant Raman reporter is embedded. Subtle aspects of their preparation can shift their limit of detection by orders of magnitude. In this protocol, we present the optimized, step-by-step procedure for generating reproducible SERRS nanoparticles with femtomolar (10−15 M) limits of detection. We provide ways of characterizing the optical properties of SERRS nanoparticles using UV/VIS and Raman spectroscopy, and their physicochemical properties using transmission electron microscopy and nanoparticle tracking analysis. We introduce several applications of these nanoprobes for biomedical research, with a focus on intraoperative cancer imaging via Raman imaging. A detailed account is provided for successful i.v. administration of SERRS nanoparticles such that delineation of cancerous lesions can be achieved in vivo and ex vivo on resected tissues without the need for specific biomarker targeting. This straightforward, yet comprehensive, protocol—from initial de novo gold nanoparticle synthesis to SERRS nanoparticle contrast-enhanced preclinical Raman imaging in animal models—takes ∼96 h.

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Figure 1: Flowchart of the workflow for SERRS-based Raman imaging of cancer in vivo.
Figure 2: Schematic representation of PEGylated SERRS nanoparticle synthesis.
Figure 3: Raman microscopy imaging setup.
Figure 4: Visual monitoring of SERRS nanoparticle synthesis.
Figure 5: SERRS nanoparticle characterization.
Figure 6: Raman spectra of two SERRS nanoparticles of different 'flavors'.
Figure 7: Contrast-enhanced Raman imaging of prostate neoplasia.

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Acknowledgements

We thank T. Shaffer (Department of Radiology, Stanford University) for verifying and reproducing our results using the described protocol. We further acknowledge the MSKCC Electron Microscopy and Molecular Cytology core facilities for technical support, and C. Andreou (MSKCC) for helpful suggestions. The following funding sources are acknowledged: NIH R01 EB017748 (M.F.K.); NIH K08 CA16396 (M.F.K.); a Pershing Square Sohn Prize by the Pershing Square Sohn Cancer Research Alliance (M.F.K.); The Dana Foundation Brain and Immuno-Imaging Grant (M.F.K.); a Dana Neuroscience Scholar Award (M.F.K.); an MSKCC Brain Tumor Center Grant (M.F.K.); an MSKCC Center for Molecular Imaging and Nanotechnology Grant (M.F.K.); an MSKCC Technology Development Grant (M.F.K.); a grant from the 'Mr. William H. and Mrs. Alice Goodwin and the Commonwealth Foundation for Cancer Research'; a grant from 'The Center for Experimental Therapeutics Center of Memorial Sloan Kettering Cancer Center' (M.F.K.); a Geoffrey Beene Cancer Research Center at MSKCC Grant and Shared Resources Award (M.F.K.); a RSNA Research Scholar Grant (M.F.K.); a Society of MSKCC Research Grant (M.F.K.); and a grant from the National Natural Science Foundation (81401461; R.H.). We also acknowledge the grant-funding support provided by the MSKCC NIH Core Grant (P30-CA008748). M.F.K. is a Damon Runyon-Rachleff Innovator supported (in part) by the Damon Runyon Cancer Research Foundation (DRR-29-14).

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Author notes

  1. Stefan Harmsen and Matthew A Wall: These authors contributed equally to this work.

  2. Department of Molecular Pharmacology, Memorial Sloan Kettering Cancer Center, New York, New York, USA.

Authors and Affiliations

  1. Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Stefan Harmsen, Matthew A Wall, Ruimin Huang & Moritz F Kircher

  2. Department of Chemistry, The Graduate Center and Hunter College, City University of New York, New York, New York, USA

    Matthew A Wall

  3. Center for Molecular Imaging and Nanotechnology (CMINT), Memorial Sloan Kettering Cancer Center, New York, New York, USA

    Moritz F Kircher

  4. Department of Radiology, Weill Cornell Medical College of Cornell University, New York, New York, USA

    Moritz F Kircher

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Contributions

S.H. and M.A.W. designed the protocol, performed the experiments and wrote the paper. R.H. performed the animal experiments and tissue processing, and wrote the paper. M.F.K. supervised the researchers and edited the paper.

Corresponding author

Correspondence to Moritz F Kircher.

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Competing interests

S.H., M.A.W. and M.F.K. have filed patent applications focused on SERRS nanoparticle composition and synthesis methods, and Raman detection hardware. M.F.K. is a cofounder of the startup company RIO Imaging, which has licensed these patents.

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Harmsen, S., Wall, M., Huang, R. et al. Cancer imaging using surface-enhanced resonance Raman scattering nanoparticles. Nat Protoc 12, 1400–1414 (2017). https://doi.org/10.1038/nprot.2017.031

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