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Spiral volumetric optoacoustic tomography for imaging whole-body biodynamics in small animals

Nature Protocols volume 18, pages 2124–2142 (2023)Cite this article

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Abstract

Fast tracking of biological dynamics across multiple murine organs using the currently commercially available whole-body preclinical imaging systems is hindered by their limited contrast, sensitivity and spatial or temporal resolution. Spiral volumetric optoacoustic tomography (SVOT) provides optical contrast, with an unprecedented level of spatial and temporal resolution, by rapidly scanning a mouse using spherical arrays, thus overcoming the current limitations in whole-body imaging. The method enables the visualization of deep-seated structures in living mammalian tissues in the near-infrared spectral window, while further providing unrivalled image quality and rich spectroscopic optical contrast. Here, we describe the detailed procedures for SVOT imaging of mice and provide specific details on how to implement a SVOT system, including component selection, system arrangement and alignment, as well as the image processing methods. The step-by-step guide for the rapid panoramic (360°) head-to-tail whole-body imaging of a mouse includes the rapid visualization of contrast agent perfusion and biodistribution. The isotropic spatial resolution possible with SVOT can reach 90 µm in 3D, while alternative steps enable whole-body scans in less than 2 s, unattainable with other preclinical imaging modalities. The method further allows the real-time (100 frames per second) imaging of biodynamics at the whole-organ level. The multiscale imaging capacity provided by SVOT can be used for visualizing rapid biodynamics, monitoring responses to treatments and stimuli, tracking perfusion, and quantifying total body accumulation and clearance dynamics of molecular agents and drugs. Depending on the imaging procedure, the protocol requires 1–2 h to complete by users trained in animal handling and biomedical imaging.

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Fig. 1: SVOT imaging system.
Fig. 2: Tomographic up-sampling.
Fig. 3: Self-gated respiratory motion rejection technique.
Fig. 4: Head-to-tail whole-body in vivo imaging of mice with SVOT.
Fig. 5: Enhancing image fidelity with a dual SOS reconstruction algorithm.
Fig. 6: Rapid overview whole-body scan and suborgan scale biodistribution mapping.
Fig. 7: High frame-rate tracking of cardiac flow dynamics using ICG.
Fig. 8: Volumetric sO2 mapping of a tumor-bearing mouse.

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Data availability

The statistical source data for Fig. 2c and Fig. 6f–h can be downloaded through the following link: https://doi.org/10.6084/m9.figshare.22257232. The raw source data for Fig. 4 can be downloaded through the following link: https://doi.org/10.6084/m9.figshare.21769340. The 3D computer aided design models of the custom-engineered animal can be downloaded through the following link: https://doi.org/10.6084/m9.figshare.21707867. All other raw datasets are available for research purposes from the corresponding author upon reasonable request. Source data are provided with this paper.

Code availability

The SVOT reconstruction code for Fig. 4a is publicly available through the following link: https://doi.org/10.6084/m9.figshare.21765299. The SVOT acquisition codes and reconstruction codes for the remaining figures are available from the corresponding author upon reasonable request.

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Acknowledgements

The authors acknowledge support from the Swiss National Science Foundation (grant 310030_192757) and the European Research Council (grant ERC-2015-CoG-682379).

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Authors and Affiliations

  1. Institute of Pharmacology and Toxicology and Institute for Biomedical Engineering, Faculty of Medicine, University of Zurich, Zurich, Switzerland

    Sandeep Kumar Kalva, Xosé Luís Deán-Ben, Michael Reiss & Daniel Razansky

  2. Institute for Biomedical Engineering, Department of Information Technology and Electrical Engineering, ETH Zurich, Zurich, Switzerland

    Sandeep Kumar Kalva, Xosé Luís Deán-Ben, Michael Reiss & Daniel Razansky

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Contributions

S.K.K., X.L.D.-B. and D.R. designed the imaging system and experiments. S.K.K. performed the experiments and analyzed the data. X.L.D.-B. provided reconstruction algorithms and assisted in data analysis. M.R. assisted in animal preparation and handling. D.R. supervised the study. All authors contributed to writing the manuscript.

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Correspondence to Daniel Razansky.

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Nature Protocols thanks Miya Ishihara, Yash Mantri and Kanyi Pu for their contribution to the peer review of this work.

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Key references using this protocol

Kalva, S. K. et al. ACS Appl. Mater. Interfaces 14, 172–178 (2022): https://doi.org/10.1021/acsami.1c17661

Ron, A. et al. Laser Phot. Rev. 15, 2000484 (2021): https://doi.org/10.1002/lpor.202000484

Deán-Ben, X. L. et al. Light Sci. Appl. 6, e16247 (2017): https://doi.org/10.1038/lsa.2016.247

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Kalva, S.K., Deán-Ben, X.L., Reiss, M. et al. Spiral volumetric optoacoustic tomography for imaging whole-body biodynamics in small animals. Nat Protoc 18, 2124–2142 (2023). https://doi.org/10.1038/s41596-023-00834-7

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