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Published October 28, 2020 | Published
Journal Article Open

Label-free imaging of lipid-rich biological tissues by mid-infrared photoacoustic microscopy


Significance: Mid-infrared (IR) imaging based on the vibrational transition of biomolecules provides good chemical-specific contrast in label-free imaging of biology tissues, making it a popular tool in both biomedical studies and clinical applications. However, the current technology typically requires thin and dried or extremely flat samples, whose complicated processing limits this technology's broader translation. Aim: To address this issue, we report mid-IR photoacoustic microscopy (PAM), which can readily work with fresh and thick tissue samples, even when they have rough surfaces. Approach: We developed a transmission-mode mid-IR PAM system employing an optical parametric oscillation laser operating in the wavelength range from 2.5 to 12 μm. Due to its high sensitivity to optical absorption and the low ultrasonic attenuation of tissue, our PAM achieved greater probing depth than Fourier transform IR spectroscopy, thus enabling imaging fresh and thick tissue samples with rough surfaces. Results: In our spectroscopy study, the CH₂ symmetric stretching at 2850  cm⁻¹ (3508 nm) was found to be an excellent source of endogenous contrast for lipids. At this wavenumber, we demonstrated label-free imaging of the lipid composition in fresh, manually cut, and unprocessed tissue sections of up to 3-mm thickness. Conclusions: Our technology requires no time-consuming sample preparation procedure and has great potential in both fast clinical histological analysis and fundamental biological studies.

Additional Information

© 2020 The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI. Received: 16 May 2020; Accepted: 14 October 2020; Published: 28 October 2020. The authors appreciate Prof. James Ballard's editing of this article. This work was supported by the National Institutes of Health under Grant Nos. R01 CA186567 (NIH Director's Transformative Research Award), R01 NS102213, U01 NS090579 (BRAIN Initiative), and U01 NS099717 (BRAIN Initiative), as well as the National Institute of Neurological Disorders and Stroke (Grant R01 NS094403). Disclosures: L.V.W. and K.M. have financial interests in Microphotoacoustics, Inc., CalPACT, LLC, and Union Photoacoustic Technologies, Ltd., which, however, did not support this work. Code, Data, and Materials Availability: The data that support the plots within this paper and other findings of this study are available from the corresponding author upon reasonable request.

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