Abstract
We present fast functional photoacoustic microscopy (PAM) for three-dimensional high-resolution, high-speed imaging of the mouse brain, complementary to other imaging modalities. We implemented a single-wavelength pulse-width-based method with a one-dimensional imaging rate of 100 kHz to image blood oxygenation with capillary-level resolution. We applied PAM to image the vascular morphology, blood oxygenation, blood flow and oxygen metabolism in both resting and stimulated states in the mouse brain.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- 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
Kim, D.S., Duong, T.Q. & Kim, S.G. Nat. Neurosci. 3, 164–169 (2000).
Sakadzić, S. et al. Nat. Methods 7, 755–759 (2010).
Hillman, E.M.C. et al. Neuroimage 35, 89–104 (2007).
Hu, S., Maslov, K., Tsytsarev, V. & Wang, L.V. J. Biomed. Opt. 14, 040503 (2009).
Deán-Ben, X.L. & Razansky, D. Light Sci. Appl. 3, e137 (2014).
Stein, E.W., Maslov, K. & Wang, L.H.V. J. Biomed. Opt. 14, 020502 (2009).
Yao, J. et al. J. Biomed. Opt. 17, 080505 (2012).
Wang, L., Maslov, K. & Wang, L.V. Proc. Natl. Acad. Sci. USA 110, 5759–5764 (2013).
Danielli, A., Favazza, C.P., Maslov, K. & Wang, L.V. Opt. Lett. 36, 769–771 (2011).
Yao, J., Maslov, K., Hu, S. & Wang, L.V. J. Biomed. Opt. 14, 054049 (2009).
Spencer, J.A. et al. Nature 508, 269–273 (2014).
Liao, L.D. et al. J. Cereb. Blood Flow Metab. 32, 938–951 (2012).
Villringer, A., Them, A., Lindauer, U., Einhäupl, K. & Dirnagl, U. Circ. Res. 75, 55–62 (1994).
Shih, A.Y. et al. J. Cereb. Blood Flow Metab. 32, 1277–1309 (2012).
Ogawa, S., Lee, T.M., Kay, A.R. & Tank, D.W. Proc. Natl. Acad. Sci. USA 87, 9868–9872 (1990).
Dunn, A.K., Devor, A., Dale, A.M. & Boas, D.A. Neuroimage 27, 279–290 (2005).
Wang, L., Xia, J., Yao, J., Maslov, K.I. & Wang, L.V. Phys. Rev. Lett. 111, 204301 (2013).
Lukianova-Hleb, E.Y., Oginsky, A.O., Olson, J.S. & Lapotko, D.O. Lasers Surg. Med. 43, 249–260 (2011).
Acknowledgements
The authors appreciate J. Ballard's close reading of the manuscript. We thank C. Li, C. Chen, W. Chapman and J. Lee for discussions and technical support. This work was sponsored by US National Institutes of Health (NIH) grants DP1 EB016986 (NIH Director's Pioneer Award), R01 CA186567 (NIH Director's Transformative Research Award), 1S10 RR028864 and R01 CA159959 (to L.V.W.), and US National Science Foundation (NSF) grant CMMI-1131758 (to J.Z.). The bright-field microscopy was performed at the Alafi Neuroimaging Laboratory of the Hope Center for Neurological Disorders, Washington University School of Medicine, which is supported by the NIH Neuroscience Blueprint Center Core grant P30 NS057105.
Author information
Authors and Affiliations
Contributions
J.Y., J.Z. and L.V.W. conceived of and designed the study. J.Y., J.-M.Y., K.I.M., L.W. and C.-H.H. constructed the imaging system. J.Y., T.T.W.W. and L.L. performed the experiments and analyzed the data. J.Y., L.W. and L.V.W. wrote the manuscript. L.V.W. supervised the whole study. All authors discussed the conceptual and practical implications of the methods.
Corresponding author
Ethics declarations
Competing interests
L.V.W. has a financial interest in Endra, Inc. and Microphotoacoustics, Inc., which, however, did not support this work. K.I.M. also has a financial interest in Microphotoacoustics, Inc.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–17, Supplementary Table 1 and Supplementary Notes 1–3 (PDF 16502 kb)
Three-dimensional rendering of mouse brain.
Three-dimensional rendering of mouse cortical vasculature imaged by PAM with an intact skull. (MOV 3838 kb)
Cross-sectional rendering of the mouse brain vasculature.
Cross-sectional rendering of the mouse brain vasculature below a 0.6×0.6 mm2 surface region, a composite image from 12 depth-scans of the optical focal zone. The shadows in deeper tissue were due to the blocking of light by superficial blood vessels. MAP, maximum amplitude projection. (MOV 1247 kb)
PAM of cerebral responses to electrical stimulations to the hindlimbs.
A 3×4 mm2 cortical area covering the somatosensory regions of both hemispheres was imaged at a volumetric rate of 1 Hz. Fractional PA amplitude changes (shown in yellow), in response to the left hindlimb stimulation (LHS) and right hindlimb stimulation (RHS), were superimposed on the vascular image (shown in red). The curve at the bottom of the video shows the average PA signal amplitude in the somatosensory regions. (MOV 2961 kb)
Volumetric rendering of oxygen saturation (sO2).
sO2 was imaged below a 3×2 mm2 surface region of a mouse brain at resting state. The sO2 values were color-coded from blue (low oxygenation) to red (high oxygenation). (MOV 4178 kb)
PAM of cerebral sO2 responses to electrical stimulations to the left hindlimb.
Left panel: sO2 map of a 2×3 mm2 cortical region covering the somatosensory region of the right hemisphere, acquired at a volumetric rate of 1 Hz. Right panel: close-up of the core responding region, marked by the dashed box in the left panel. The curve at the right bottom of the video shows the average sO2 in the close-up region. Note that the resting-state frames acquired between 120 seconds and 200 seconds are omitted in the video. (MOV 1727 kb)
Rights and permissions
About this article
Cite this article
Yao, J., Wang, L., Yang, JM. et al. High-speed label-free functional photoacoustic microscopy of mouse brain in action. Nat Methods 12, 407–410 (2015). https://doi.org/10.1038/nmeth.3336
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nmeth.3336
This article is cited by
-
Free-moving-state microscopic imaging of cerebral oxygenation and hemodynamics with a photoacoustic fiberscope
Light: Science & Applications (2024)
-
Cortex-wide transcranial localization microscopy with fluorescently labeled red blood cells
Nature Communications (2024)
-
Functional photoacoustic imaging: from nano- and micro- to macro-scale
Nano Convergence (2023)
-
Photoacoustic vector tomography for deep haemodynamic imaging
Nature Biomedical Engineering (2023)
-
Dynamic 3D imaging of cerebral blood flow in awake mice using self-supervised-learning-enhanced optical coherence Doppler tomography
Communications Biology (2023)