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Millimetre-deep micrometre-resolution vibrational imaging by shortwave infrared photothermal microscopy

Nature Photonics volume 18, pages 944–951 (2024)Cite this article

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Abstract

Deep tissue chemical imaging has a vital role in biological and medical applications. Current approaches suffer from water absorption and tissue scattering, which limits imaging depth to hundreds of micrometres. The shortwave infrared spectral window allows deep tissue imaging but typically features unsatisfactory spatial resolution or low detection sensitivity. Here we present a shortwave infrared photothermal (SWIP) microscope for millimetre-deep vibrational imaging with micrometre lateral resolution. By pumping the overtone transition of carbon–hydrogen bonds and probing the subsequent photothermal lens with shortwave infrared light, SWIP can obtain chemical contrast from 1 μm polymer particles located at 800 μm depth in a highly scattering phantom. The amplitude of the SWIP signal is shown to be 63 times larger than that of the optically probed photoacoustic signal. We further demonstrate that SWIP can resolve intracellular lipids across an intact tumour spheroid and the layered structure in thick liver, skin, brain and breast tissues. SWIP microscopy fills a gap in vibrational imaging with subcellular resolution and millimetre-level penetration, which heralds broad potential for life science and clinical applications.

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Fig. 1: SWIP microscope principle and schematic.
Fig. 2: Comparison of optically detected PT and PA signals.
Fig. 3: SWIP microscope performance.
Fig. 4: SWIP imaging of cancer cells and spheroids.
Fig. 5: SWIP imaging of lipids in biological tissues.
Fig. 6: Penetration depth versus spatial resolution of vibrational imaging modalities.

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Data availability

Source data are available via figshare at https://doi.org/10.6084/m9.figshare.25687185 (ref. 47).

Code availability

Code is available via figshare at https://doi.org/10.6084/m9.figshare.25687185 (ref. 47).

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Acknowledgements

Human breast samples from the Susan G. Komen Tissue Bank at the IU Simon Cancer Center were used in this study. This work is supported by NIH R35GM136223, R33CA261726, R01 HL125385, R01 EB032391 and R01CA224275 to J.-X.C. We thank L. Lu for advice on the photothermal detection scheme and G. Chen for help in understanding photoacoustic signal propagation. We acknowledge M. Cherepashensky for proofreading of the paper.

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Author notes

  1. These authors contributed equally: Hongli Ni, Yuhao Yuan.

Authors and Affiliations

  1. Department of Electrical and Computer Engineering, Boston University, Boston, MA, USA

    Hongli Ni, Yuhao Yuan, Mingsheng Li, Xiaowei Ge, Jiaze Yin, Le Wang & Ji-Xin Cheng

  2. Department of Chemistry, Boston University, Boston, MA, USA

    Yifan Zhu & Ji-Xin Cheng

  3. Department of Biomedical Engineering, Boston University, Boston, MA, USA

    Chinmayee Prabhu Dessai & Ji-Xin Cheng

  4. Photonics Center, Boston University, Boston, MA, USA

    Ji-Xin Cheng

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Contributions

H.N. developed the SWIP system, designed the experiments, processed the data, built the theoretical models and drafted the paper. Y.Y. helped in the experiments and paper writing, performed the theoretical analysis, and prepared cancer cell and spheroid samples. M.L. provided the mouse tissue and helped in data analysis. Y.Z. contributed to the project formulation and building of theoretical models. X.G. contributed to the project formulation and data analysis. J.Y. helped in optimizing the photothermal detection scheme. C.P.D. prepared the human breast biopsy sample. L.W. helped in the SRS experiment. J.-X.C. initialized the project, revised the paper and provided scientific guidance.

Corresponding author

Correspondence to Ji-Xin Cheng.

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Competing interests

J.-X.C. declares financial interest with Photothermal Spectroscopy Corp, which did not support this work. The other authors declare no competing interests.

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Peer review information

Nature Photonics thanks Keisuke Goda and Lu Wei for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–12, Texts 1–6, Tables 1 and 2, and captions for Videos 1 and 2.

Supplementary Video 1

Layer-by-layer view of volumetric SWIP imaging of an OVCAR-5-cisR spheroid. Field of view, 200 × 200 μm.

Supplementary Video 2

Three-dimensional rotation view of volumetric SWIP imaging of an OVCAR-5-cisR spheroid.

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Ni, H., Yuan, Y., Li, M. et al. Millimetre-deep micrometre-resolution vibrational imaging by shortwave infrared photothermal microscopy. Nat. Photon. 18, 944–951 (2024). https://doi.org/10.1038/s41566-024-01463-6

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