Optical-Resolution Photoacoustic Microscopy for Volumetric and Spectral Analysis of Histological and Immunochemical Samples

Authors

  • Dr. Yu Shrike Zhang,

    1. The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332 (USA)
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    • These authors contributed equally to this work.

  • Dr. Junjie Yao,

    1. Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)
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    • These authors contributed equally to this work.

  • Chi Zhang,

    1. Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)
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  • Lei Li,

    1. Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)
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  • Prof. Lihong V. Wang,

    Corresponding author
    1. Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)
    • Lihong V. Wang, Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)===

      Younan Xia, The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332 (USA)===

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  • Prof. Younan Xia

    Corresponding author
    1. The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332 (USA)
    2. School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332 (USA)
    • Lihong V. Wang, Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130 (USA)===

      Younan Xia, The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332 (USA)===

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  • This work was supported by NIH grants DP1 OD000798 (NIH Director’s Pioneer Award) to Y.X., and DP1 EB016986 (NIH Director’s Pioneer Award), R01 CA186567 (NIH Director’s Transformative Research Award), and R01 CA159959 to L.V.W. L.V.W. has a financial interest in Endra, Inc., and Microphotoacoustics, Inc., which did not support this work. Part of the research was performed at the Alafi Neuroimaging Laboratory, the Hope Center for Neurological Disorders in Washington University School of Medicine, which is supported by the NIH Neuroscience Blueprint Center Core Grant P30 NS057105.

Abstract

Optical-resolution photoacoustic microscopy (OR-PAM) is an imaging modality with superb penetration depth and excellent absorption contrast. Here we demonstrate, for the first time, that this technique can advance quantitative analysis of conventional chromogenic histochemistry. Because OR-PAM can quantify the absorption contrast at different wavelengths, it is feasible to spectrally resolve the specific biomolecules involved in a staining color. Furthermore, the tomographic capability of OR-PAM allows for noninvasive volumetric imaging of a thick sample without microtoming it. By immunostaining the sample with different chromogenic agents, we further demonstrated the ability of OR-PAM to resolve different types of cells in a coculture sample with imaging depths up to 1 mm. Taken together, the integration of OR-PAM with (immuno)histochemistry offers a simple and versatile technique with broad applications in cell biology, pathology, tissue engineering, and related biomedical studies.

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