Theranostics 2014; 4(6):604-613. doi:10.7150/thno.7996

Research Paper

Real-Time Assessment of Tissue Hypoxia In Vivo with Combined Photoacoustics and High-Frequency Ultrasound

Marco Gerling1*, Ying Zhao2,3*, Salvatore Nania3, K. Jessica Norberg3, Caroline S. Verbeke4, Benjamin Englert1, Raoul V. Kuiper6, Åsa Bergström1, Moustapha Hassan5, Albrecht Neesse7, J. Matthias Löhr1,3, Rainer L. Heuchel1,3 ✉

1. Center of Biosciences, Department of Biosciences and Nutrition, Karolinska Institutet, Stockholm, Sweden;
2. StratCan Preclinical Cancer Test Facility, Karolinska Institutet, Stockholm, Sweden;
3. Pancreas Research Laboratory, CLINTEC, Karolinska Institutet, Stockholm, Sweden;
4. Division of Pathology, Karolinska Institutet, Stockholm, Sweden;
5. Clinical Research Centre, Karolinska University Hospital-Huddinge, Stockholm, Sweden;
6. Core Facility for Morphologic Phenotype Analysis, Clinical Research Center, Karolinska Institutet, Stockholm, Sweden;
7. Department of Gastroenterology, Endocrinology and Metabolism, Philipps University Marburg, Marburg, Germany.
* equal contribution.


Purpose: In preclinical cancer studies, non-invasive functional imaging has become an important tool to assess tumor development and therapeutic effects. Tumor hypoxia is closely associated with tumor aggressiveness and is therefore a key parameter to be monitored. Recently, photoacoustic (PA) imaging with inherently co-registered high-frequency ultrasound (US) has reached preclinical applicability, allowing parallel collection of anatomical and functional information. Dual-wavelength PA imaging can be used to quantify tissue oxygen saturation based on the absorbance spectrum differences between hemoglobin and deoxyhemoglobin.

Experimental Design: A new bi-modal PA/US system for small animal imaging was employed to test feasibility and reliability of dual-wavelength PA for measuring relative tissue oxygenation. Murine models of pancreatic and colon cancer were imaged, and differences in tissue oxygenation were compared to immunohistochemistry for hypoxia in the corresponding tissue regions.

Results: Functional studies proved feasibility and reliability of oxygenation detection in murine tissue in vivo. Tumor models exhibited different levels of hypoxia in localized regions, which positively correlated with immunohistochemical staining for hypoxia. Contrast-enhanced imaging yielded complementary information on tissue perfusion using the same system.

Conclusion: Bimodal PA/US imaging can be utilized to reliably detect hypoxic tumor regions in murine tumor models, thus providing the possibility to collect anatomical and functional information on tumor growth and treatment response live in longitudinal preclinical studies.

Keywords: Photoacoustics, High-Frequency Ultrasound, Hypoxia.

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How to cite this article:
Gerling M, Zhao Y, Nania S, Norberg KJ, Verbeke CS, Englert B, Kuiper RV, Bergström Å, Hassan M, Neesse A, Löhr JM, Heuchel RL. Real-Time Assessment of Tissue Hypoxia In Vivo with Combined Photoacoustics and High-Frequency Ultrasound. Theranostics 2014; 4(6):604-613. doi:10.7150/thno.7996. Available from