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High-speed molecular spectral imaging of tissue with stimulated Raman scattering


To date, medical imaging of tissues has largely relied on time-consuming staining processes, and there is a need for rapid, label-free imaging techniques. Stimulated Raman scattering microscopy offers a three-dimensional, real-time imaging capability with chemical specificity. However, it can be difficult to differentiate between several constituents in tissues because their spectral characteristics can overlap. Furthermore, imaging speeds in previous multispectral stimulated Raman scattering imaging techniques were limited. Here, we demonstrate label-free imaging of tissues by 30 frames/s stimulated Raman scattering microscopy with frame-by-frame wavelength tunability. To produce multicolour images showing different constituents, spectral images were processed by modified independent component analysis, which can extract small differences in spectral features. We present various imaging modalities such as two-dimensional spectral imaging of rat liver, two-colour three-dimensional imaging of a vessel in rat liver, spectral imaging of several sections of intestinal villi in mouse, and in vivo spectral imaging of mouse ear skin.

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Figure 1: High-speed SRS spectral microscopy and modified ICA.
Figure 2: Spectral imaging of rat liver tissue.
Figure 3: Three-dimensional, two-colour imaging of a vessel in rat liver.
Figure 4: Sectioned spectral imaging of intestinal villi in the mouse.
Figure 5: In vivo spectral imaging of the sebaceous gland of mouse ear skin.
Figure 6: Spectral imaging in fingerprint region.

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The authors thank H. Watanabe for his kind support with the experiment on mouse skin imaging. They also thank N. Smith for his careful editing of this manuscript. This research was supported by Japan Science and Technology (JST)–Precursory Research for Embryonic Science and Technology (PRESTO) (Y. Ozeki) and partially supported by KAKENHI (21248040).

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Y. Ozeki conceived the idea and drafted the manuscript. Y. Ozeki and W.U. built the instrument and coded data analysis programs. K.S. and N.N. built and modified the fibre laser. Y. Otsuka and S.S. prepared biological samples. Y. Ozeki, W.U. and Y. Otsuka conducted the experiments. H.H., K.F. and K.I. supervised the research.

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Correspondence to Yasuyuki Ozeki.

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The authors declare no competing financial interests.

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Ozeki, Y., Umemura, W., Otsuka, Y. et al. High-speed molecular spectral imaging of tissue with stimulated Raman scattering. Nature Photon 6, 845–851 (2012).

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