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. Author manuscript; available in PMC: 2022 Apr 15.
Published in final edited form as: ACS Nano. 2021 Nov 19;15(12):19956–19969. doi: 10.1021/acsnano.1c07470

Figure 5.

Figure 5.

Correlation of the five-color NIR-SERRS nanoparticles injection ratio with the multiplexed Raman images of the tumors. (a–c) Tumor-xenograft nude mice (left, upper panel), and the five-color multiplexed Raman images of their tumors (right, upper panel), for which the ROI was defined as rectangles in the photograph. The multiplexed images were generated through the spectral unmixing of Raman spectra in each pixel, followed by color-coding (lower panels) and merge (right, upper panel). (d–f) Representative Raman spectra (upper panel) and the normalized concentrations (ratios) of the five-color NIR-SERRS nanoparticles (lower panel) in the tumors, obtained from the spectral unmixing of the Raman spectra in each pixel and averaged over the entire images of (a) (d), (b) (e), and (c) (f). In each pixel, the five-color nanoparticle concentrations were normalized to the concentration of the IR813-coded (d) or IR797-coded NIR-SERRS nanoparticles (e and f) that were set as 5 (d and e) or 10 (f). The error bars: the standard deviations of the spectrally unmixed concentrations calculated from all the pixelated Raman spectra in the ROIs. (g–i) Ex vivo tumor tissues (left, upper panel) and their respective five-color multiplexed Raman images (right, upper panel). The multiplexed images were generated through the spectral unmixing of Raman spectra in each pixel, followed by color-coding (lower panels) and merge (right, upper panel). The color-coded Raman images were organized with the order of increasing brightness from the left to right (lower panel, a–c, and g–i). The mixture of 1:2:3:4:5 molar ratio of IR780, DTTC, HDITC, IR797, and IR813-coded nanoparticles (mouse #1, (a), (d), and (g)), respectively, or 1:2:3:4:5 molar ratio of DTTC, IR780, HDITC, IR813, IR797 (mouse #2, (b), (e), and (h)), or 1:2:4:8:10 molar ratio of DTTC, IR780, HDITC, IR813, IR797 (mouse #3, (c), (f), and (i)), respectively, was prepared and concurrently injected via tail-vein. Color codes of the five-color NIR-SERRS nanoparticles: DTTC, magenta-purple; HDITC, blue; IR780, orange; IR797, red; and IR813, green. Scale bar: 10 mm.