Abstract
Although optical absorption is strongly associated with the physiological status of biological tissue, existing high-resolution optical imaging modalities, including confocal microscopy1,2, two-photon microscopy3,4 and optical coherence tomography5, do not sense optical absorption directly. Furthermore, optical scattering prevents these methods from imaging deeper than ∼1 mm below the tissue surface. Here we report functional photoacoustic microscopy (fPAM), which provides multiwavelength imaging of optical absorption and permits high spatial resolution beyond this depth limit with a ratio of maximum imaging depth to depth resolution greater than 100. Reflection mode, rather than orthogonal or transmission mode, is adopted because it is applicable to more anatomical sites than the others. fPAM is demonstrated with in vivo imaging of angiogenesis, melanoma, hemoglobin oxygen saturation (sO2) of single vessels in animals and total hemoglobin concentration in humans.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
£139.00 per year
only £11.58 per issue
Buy this article
- Purchase on SpringerLink
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Wilson, T. & Sheppard, C. Theory and Practice of Scanning Optical Microscopy (Academic Press, London, 1984).
Sipkins, D.A. et al. In vivo imaging of specialized bone marrow endothelial microdomains for tumour engraftment. Nature 435, 969–973 (2005).
Denk, W., Strickler, J.H. & Webb, W.W. Two-photon laser scanning fluorescence microscopy. Science 248, 73–76 (1990).
So, P.T.C., Dong, C.Y., Masters, B.R. & Berland, K.M. Two-photon excitation fluorescence microcopy. Annu. Rev. Biomed. Eng. 2, 399–429 (2000).
Huang, D. et al. Optical coherence tomography. Science 254, 1178–1181 (1991).
Sun, T. & Diebold, G.J. Generation of ultrasonic waves from a layered photoacoustic source. Nature 355, 806–808 (1992).
Duck, F.A. . Physical Properties of Tissue (Academic Press London, 1990).
Wang, X. et al. Noninvasive laser-induced photoacoustic tomography for structural and functional in vivo imaging of the brain. Nat. Biotechnol. 21, 803–806 (2003).
Hoelen, C.G.A., de Mul, F.F.M., Pongers, R. & Dekker, A. Three-dimensional photoacoustic imaging of blood vessels in tissue. Opt. Lett. 23, 648–650 (1998).
Oraevsky, A.A. & Karabutov, A.A. Optoacoustic tomography. in Biomedical Photonics Handbook, vol. PM125 (ed. Vo-Dinh, T.) (CRC Press, Boca Raton, Florida, 2003).
Briggs, G.A.D. Acoustic Microscopy, p. 27 (Clarendon, Oxford, 1992).
Maslov, K., Stoica, G. & Wang, L.V. In vivo dark-field reflection-mode photoacoustic microscopy. Opt. Lett. 30, 625–627 (2005).
Carmeliet, P. & Jain, R.K. Angiogenesis in cancer and other diseases. Nature 407, 249–257 (2000).
Jobsis, F.F. Noninvasive, infrared monitoring of cerebral and myocardial oxygen sufficiency and circulatory parameters. Science 198, 1264–1267 (1977).
Chance, B. et al. Comparison of time-resolved and -unresolved measurements of deoxyhemoglobin in brain. Proc. Natl. Acad. Sci. USA 85, 4971–4975 (1988).
Tsao, M.U., Sethna, S.S., Sloan, C.H. & Wyngarden, L.J. Spectrophotometric determination of the oxygen saturation of whole blood. J. Biol. Chem. 217, 479–488 (1955).
Vanzetta, I. & Grinvald, A. Increased cortical oxidative metabolism due to sensory stimulation: implications for functional brain imaging. Science 286, 1555–1558 (1999).
Levasseur, J., Wei, E., Raper, A., Kontos, H. & Patterson, J. Detailed description of a cranial window technique for acute and chronic experiments. Stroke 6, 308–317 (1975).
Hall, D.G. & Stoica, G. Characterization of brain and bone-metastasizing clones selected from an ethylnitrosourea-induced rat mammary carcinoma. Clin. Exp. Metastasis 12, 283–295 (1994).
Ambach, G. & Palkovits, M. Blood supply of the rat hypothalamus I. nucleus supraopticus. Acta Morphol. Acad. Sci. Hung. 22, 291–310 (1974).
Acknowledgements
We thank O. Craciun, J. Oh, G. Ku, M.L. Li and G. Lungu for experimental assistance. This work was sponsored by National Institutes of Health grants R01 EB000712 and R01 NS46214.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Fig. 1
An illustrative example showing the penetration depth of fPAM in tissue. (PDF 69 kb)
Supplementary Fig. 2
Images of vasculature in a rat acquired in vivo by fPAM at the isosbestic optical wavelength of 584 nm before, two days post, and five days post subcutaneous inoculation of BR7C5 tumor cells. (PDF 76 kb)
Supplementary Table 1
Comparison among the modern high-resolution microscopic imaging techniques, whose depth-to-resolution ratios are all greater than 100. (PDF 15 kb)
Supplementary Video 1
(1.7 MB) Movie for the composite volumetric visualization of a melanoma in the skin acquired in vivo. (AVI 1774 kb)
Rights and permissions
About this article
Cite this article
Zhang, H., Maslov, K., Stoica, G. et al. Functional photoacoustic microscopy for high-resolution and noninvasive in vivo imaging. Nat Biotechnol 24, 848–851 (2006). https://doi.org/10.1038/nbt1220
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nbt1220
This article is cited by
-
Functional photoacoustic imaging: from nano- and micro- to macro-scale
Nano Convergence (2023)
-
Advances in photoacoustic imaging aided by nano contrast agents: special focus on role of lymphatic system imaging for cancer theranostics
Journal of Nanobiotechnology (2023)
-
Label-free biomedical optical imaging
Nature Photonics (2023)
-
All-optical optoacoustic micro-tomography in reflection mode
Biomedical Engineering Letters (2023)
-
Photoacoustic Microscopic Imaging of Cerebral Vessels for Intensive Monitoring of Metabolic Acidosis
Molecular Imaging and Biology (2023)