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Optical-resolution photoacoustic microscopy with a needle-shaped beam

Abstract

Optical-resolution photoacoustic microscopy can visualize wavelength-dependent optical absorption at the cellular level. However, this technique suffers from a limited depth of field due to the tight focus of the optical excitation beam, making it challenging to acquire high-resolution images of samples with uneven surfaces or high-quality volumetric images without z scanning. To overcome this limitation, we propose needle-shaped beam photoacoustic microscopy, which can extend the depth of field to around a 28-fold Rayleigh length via customized diffractive optical elements. These diffractive optical elements generate a needle-shaped beam with a well-maintained beam diameter, a uniform axial intensity distribution and negligible sidelobes. The advantage of using needle-shaped beam photoacoustic microscopy is demonstrated via both histology-like imaging of fresh slide-free organs using a 266 nm laser and in vivo mouse-brain vasculature imaging using a 532 nm laser. This approach provides new perspectives for slide-free intraoperative pathological imaging and in vivo organ-level imaging.

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Fig. 1: Principle of NB-PAM with a customized DOE.
Fig. 2: Characterization of UV-NB-PAM in comparison with that of conventional UV-GB-PAM.
Fig. 3: Volumetric imaging of carbon particles using UV-GB-PAM and UV-NB-PAM with a 200 × 1.2 μm NB.
Fig. 4: Depth-resolved imaging of carbon fibres for VIS-GB-PAM and for VIS-NB-PAM with the 1,000 × 2.3 μm NB.
Fig. 5: Label-free UV-GB-PAM and UV-NB-PAM with the 200 × 1.2 μm NB for slide-free fresh mouse lung and brain samples.
Fig. 6: In vivo label-free depth-encoded VIS-NB-PAM with the 1,000 × 2.3 μm NB and VIS-GB-PAM of brain vasculature with and without a skull.

Data availability

The data that support the findings of this study are available within the paper and its Supplementary Information. The raw data are too large to be publicly shared, yet they are available for research purposes from the corresponding authors upon reasonable request.

Code availability

The code that supports the plots and images within this paper is available from the corresponding author upon reasonable request.

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Acknowledgements

L.V.W. was sponsored by the United States National Institutes of Health grants R01 EB028277, U01 NS099717 (BRAIN Initiative) and R35 CA220436 (Outstanding Investigator Award). A.d.l.Z. was supported by the National Institutes of Health grants DP50D012179 and K23CA211793, the United States National Science Foundation (NSF 1438340) and the United States Air Force (FA9550–15–1–0007).

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Authors

Contributions

R.C. and L.V.W. designed the experiment. R.C., L.L. and Y.Z. built the PAM system. J.Z. designed and fabricated the DOEs. L.D. contributed to the mask preparation and wafer dividing. L.J. and Q.Z. manufactured the ultrasonic transducer. R.C. prepared the sample and animals and performed the imaging experiment. R.C., S.D. and Y.L. contributed to image processing. L.V.W. and A.d.l.Z. supervised the project. All authors were involved in discussions and manuscript preparation.

Corresponding authors

Correspondence to Adam de la Zerda or Lihong V. Wang.

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Competing interests

L.V.W. has a financial interest in MicroPhotoAcoustics, CalPACT and Union Photoacoustic Technologies, although they did not support this work. The remaining authors declare no competing interests.

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Nature Photonics thanks Ruiqing Ni and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Note, Figs. 1–9 and Tables 1–3.

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Cao, R., Zhao, J., Li, L. et al. Optical-resolution photoacoustic microscopy with a needle-shaped beam. Nat. Photon. 17, 89–95 (2023). https://doi.org/10.1038/s41566-022-01112-w

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