Research ArticleCancer Imaging

Surface-enhanced resonance Raman scattering nanostars for high-precision cancer imaging

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Science Translational Medicine  21 Jan 2015:
Vol. 7, Issue 271, pp. 271ra7
DOI: 10.1126/scitranslmed.3010633
  • Fig. 1. Characterization of SERRS nanostars.

    (A) Schematic and three-dimensional representations of the SERRS nanostar geometry. Transmission electron micrographs of a single SERRS nanostar and a population of SERRS nanostars are shown. (B) SERRS nanostar size distribution as determined by nanoparticle tracking analysis. (C) Raman spectra showing photostability of 1 nM SERRS nanostars during continuous irradiation (100 mW laser power) for 30 min. Spectra were acquired at 5-min intervals (50-μW laser power, 1-s acquisition time, 5× objective). cps, counts/s. (D) Limit of detection of SERRS nanostars in solution was 1.5 fM at 100 mW, 1.5-s acquisition time, 5× objective. Data are representative of three separate experiments. (E and F) Serum stability of the SERRS signal intensity (E) and hydrodynamic diameter (F) of 1.0 nM PEGylated SERRS nanostars during incubation in 50% mouse serum. Data are means ± SEM (n = 3).

  • Fig. 2. Imaging of breast cancer in the MMTV-PyMT mouse model.

    Images are representative of n = 6 mice. (A and B) Two adjacent tumors developed in the upper and lower right thoracic mammary glands. Gray dashed box in photograph indicates areas scanned with Raman imaging. After imaging, the first tumor was resected along the white dotted line (A). Anti-PEG IHC staining shows presence of SERRS nanostars in the tumor. The second tumor was then also resected along the white dotted line (B). (C) Gray dashed box in photograph indicates resection bed after removal of tumors in (A) and (B). Staining for PyMT indicated residual microscopic tumor (arrow). Raman signal intensity is displayed in counts/s. Insets in the α-PEG images are 2× magnification views.

  • Fig. 3. Imaging microscopic tumor infiltration into the skin in the Ink4a/Arf−/− fibrosarcoma model.

    Images are representative of n = 4 mice. (A) White dashed box in photograph highlights the primary tumor on the right shoulder of an Ink4a/Arf−/− fibrosarcoma-bearing mouse after hair removal. Despite the red discoloration, the skin overlying the tumor is intact. Images were obtained before surgical exposure of the tumor. (B) Photograph on the upper left shows the bulk tumor (black box 1) after the overlying skin (gray box 2) had been lifted off. Raman images of each boxed area were acquired, focusing on the bulk tumor (box 1) and the skin overlying the tumor (box 2), respectively. (C and D) Histological analysis of the resected bulk tumor (C) and the skin overlying the tumor (D) at different magnifications of indicated regions (boxes). Antibody against the marker Ki-67 (α-MKI67) indicated cell proliferation and α-PEG stained for SERRS nanostars. Raman signal intensity is displayed in counts/s.

  • Fig. 4. Microscopic infiltration at tumor margins and regional satellite metastases in the human DDLS mouse model.

    Images are representative of n = 7 mice. (A) SERRS nanostars were detected by Raman imaging of the bulk tumor. IHC staining for human vimentin indicated the presence of tumor cells; anti-PEG IHC staining indicated the presence of SERRS nanostars. (B) Raman image of the resection bed acquired after surgical excision of the bulk tumor in (A); resection was guided by white light only. IHC images on the far right are magnified views of the areas indicated with arrows 1 and 2. (C) In a different mouse bearing a liposarcoma, multiple small foci of Raman signal (arrows 1 to 5) were found ~10 mm away from the margins of the bulk tumor. As confirmed by IHC, each of these five SERRS nanostar–positive foci correlated with a separate tumor cell cluster (vimentin+) as small as 100 μm (micrometastases). Images on far right are magnified views of the metastases labeled 4 and 5. Raman signal intensity is displayed in counts/s.

  • Fig. 5. Imaging of PDAC and pancreatic intraepithelial lesion in the KPC mouse model.

    Images are representative of n = 5 mice. (A) In situ photograph of the exposed upper abdomen in a mouse with a PDAC in the head of the pancreas (outlined with white dotted line). Corresponding Raman images, showing SERRS nanostar signal in the macroscopically visible tumor in the head as well as small scattered foci of SERRS signal in other normal-appearing regions of the pancreas, are also shown. (B) Photographic and high-resolution Raman images of the excised pancreas from (A). (C) H&E staining of the whole pancreas, including PDAC (arrow 1) and PanIN (arrow 2). Histology and keratin 19 (KRT19) staining in regions 1 and 2 confirmed lesions. Insets are 4× magnification views. Raman signal intensity is displayed in counts/s.

  • Fig. 6. Imaging different stages and grades of prostatic neoplasia within the same prostate in the Hi-Myc mouse model.

    Images are representative of n = 5 mice. (A) Sequential resection of the prostatic tumors with corresponding Raman images. White dotted lines indicate the margins of each resection. (B) Histological staining for the tumor marker MYC, androgen receptor (AR), and PEG (indicating the presence of SERRS nanostars; insets are 2× magnification views) of the respective resected tumors in (A). Raman signal intensity is displayed in counts/s. PIN, prostatic intraepithelial neoplasia.

  • Fig. 7. Macropinocytosis is a major contributor to SERRS nanostar uptake by tumor cells.

    Four small molecule inhibitors—EIPA, NVP-BEZ235, wortmannin, and cytochalasin D (Cyto D)—were applied in vitro to tumor cell lines established from primary spontaneous tumors of the MMTV-PyMT [AT-3], KPC [PCC-9], and Hi-Myc [Myc-CaP] transgenic mice and DDLS8817 liposarcoma cells. Raman images of the cells were acquired, and the Raman signal from the accumulated SERRS nanostars was quantified and normalized to the cell number. Data are means ± SD normalized to dimethyl sulfoxide (DMSO) vehicle control (defined as 100%) and are representative of three separate experiments. *P < 0.05 versus the DMSO control; unpaired t-test.

Supplementary Materials

  • Supplementary Material for:

    Surface-enhanced resonance Raman scattering nanostars for highprecision cancer imaging

    Stefan Harmsen, Ruimin Huang, Matthew A. Wall, Hazem Karabeber, Jason M. Samii, Massimiliano Spaliviero, Julie R. White, Sébastien Monette, Rachael O'Connor, Kenneth L. Pitter, Stephen A. Sastra, Michael Saborowski, Eric C. Holland, Samuel Singer, Kenneth P. Olive, Scott W. Lowe, Ronald G. Blasberg, Moritz F. Kircher*

    *Corresponding author. E-mail: kircherm{at}mskcc.org

    Published 21 January 2015, Sci. Transl. Med. 7, 271ra7 (2015)
    DOI: 10.1126/scitranslmed.3010633

    This PDF file includes:

    • Methods
      Fig. S1. Characterization of SERRS nanostars.
    • Fig. S2. Tissue distribution and biliary excretion of SERRS nanostars.
    • Fig. S3. Potential clinical applications of SERRS nanostars.
    • Table S1. Zeta potentials of nanostars.
    • References (43, 44)

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