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High-speed coherent Raman fingerprint imaging of biological tissues

Nature Photonics volume 8, pages 627–634 (2014)Cite this article

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Abstract

An imaging platform based on broadband coherent anti-Stokes Raman scattering has been developed that provides an advantageous combination of speed, sensitivity and spectral breadth. The system utilizes a configuration of laser sources that probes the entire biologically relevant Raman window (500–3,500 cm–1) with high resolution (<10 cm–1). It strongly and efficiently stimulates Raman transitions within the typically weak ‘fingerprint’ region using intrapulse three-colour excitation, and utilizes the non-resonant background to heterodyne-amplify weak Raman signals. We demonstrate high-speed chemical imaging in two- and three-dimensional views of healthy murine liver and pancreas tissues as well as interfaces between xenograft brain tumours and the surrounding healthy brain matter.

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Figure 1: Coherent Raman imaging with BCARS microspectroscopy.
Figure 2: CRI of murine liver tissue.
Figure 3: Three-dimensional CRI of murine pancreatic ducts.
Figure 4: Histopathology using broadband CRI.

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Acknowledgements

The authors thank Q. Wu, J. Hale and M. Sinyuk for preparing the pathological tissue specimens and S. Miller for preparation of neat chemical specimens. C.H.C., J.M.H. and C.M.H. also thank the National Research Council for support through the Research Associate Program (RAP). This work was supported in part by NIH/NIBIB grant 2P41EB001046-11.

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Authors and Affiliations

  1. Biosystems and Biomaterials Division, National Institute of Standards and Technology, Gaithersburg, 20899, Maryland, USA

    Charles H. Camp Jr, Young Jong Lee, Christopher M. Hartshorn & Marcus T. Cicerone

  2. Semiconductor and Dimensional Metrology Division, National Institute of Standards and Technology, Gaithersburg, 20899, Maryland, USA

    John M. Heddleston & Angela R. Hight Walker

  3. Department of Stem Cell Biology and Regenerative Medicine, Cleveland Clinic, Cleveland, 44195, Ohio, USA

    Jeremy N. Rich

  4. Department of Cellular and Molecular Medicine, Cleveland Clinic, Cleveland, 44195, Ohio, USA

    Justin D. Lathia

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  1. Charles H. Camp Jr
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Contributions

C.H.C. performed all experiments, analysed all data and drafted the manuscript. M.T.C. and C.H.C. designed all experiments and constructed the final manuscript. M.T.C. and Y.J.L. conceptualized the complementary two/three-colour excitation scheme. C.H.C. constructed the instrument, modified the laser system and developed the high-speed acquisition and processing software. C.H.C., Y.J.L., C.M.H. and M.T.C. developed the signal-processing methodology and protocols. M.T.C. developed the Kramers–Kronig transform and C.H.C. developed the parallelized, high-speed implementation. A.R.H.W., J.M.H., J.N.R. and J.D.L. provided materials and/or the tumour sections and provided histopathology insights and direction. J.M.H. assisted in performing the tumour section study, as well as contributing to the text of the manuscript. A.R.H.W., J.M.H. and C.H.C. collected the spontaneous Raman spectra of glycerol and C.H.C. performed the analysis. C.H.C. developed the presented mathematical framework of CARS generation and associated efficiencies with two/three-colour stimulation. M.T.C. supervised the study.

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Correspondence to Marcus T. Cicerone.

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Camp Jr, C., Lee, Y., Heddleston, J. et al. High-speed coherent Raman fingerprint imaging of biological tissues. Nature Photon 8, 627–634 (2014). https://doi.org/10.1038/nphoton.2014.145

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