Super-resolution functional photoacoustic microscopy via label-free cell tracking

Fenghe Zhong; Zhuoying Wang; Youngseop Lee; Jiaxiao Han; Naidi Sun; Shuo Yang; Shengyun Ji; Hao F. Zhang; Cheng Sun; Song Hu

doi:10.1038/s41377-026-02235-3

Your Location：

Home >

Browse articles >

Super-resolution functional photoacoustic microscopy via label-free cell tracking

Original Articles

- Super-resolution functional photoacoustic microscopy via label-free cell tracking
- Light: Science & Applications Vol. 15, Issue 5, Pages: 1581-1593(2026)
- Affiliations：
  
  1.Department of Biomedical Engineering, Washington University in St. Louis, St. Louis, MO 63130, USA
  2.Department of Biomedical Engineering, Northwestern University, Evanston, IL 60208, USA
  3.Department of Mechanical Engineering, Northwestern University, Evanston, IL 60208, USA
- Author bio：
  
  Song Hu (songhu@wustl.edu)
- Funds：
- DOI：10.1038/s41377-026-02235-3
  CLC：
- Received：13 October 2025，
  
  Revised：2026-02-02，
  
  Accepted：08 February 2026，
  
  Online First：03 March 2026，
  
  Published：31 May 2026
- Accepted：
Scan QR Code
Zhong, F. H. et al. Super-resolution functional photoacoustic microscopy via label-free cell tracking. Light: Science & Applications, 15, 1581-1593 (2026).
DOI：

Zhong, F. H. et al. Super-resolution functional photoacoustic microscopy via label-free cell tracking. Light: Science & Applications, 15, 1581-1593 (2026). DOI： 10.1038/s41377-026-02235-3.

摘要

Abstract

Microvascular function and oxygen metabolism are central to tissue and organ health. However

label-free methods for imaging oxygen dynamics in three-dimensional (3D) microvascular networks at the level of single red blood cells (RBCs)—the fundamental units of oxygen transport in vivo—remain lacking. Here

we introduce super-resolution functional photoacoustic microscopy (SR-fPAM)

which spatiotemporally tracks RBC movements under dual-wavelength excitation. SR-fPAM reconstructs super-resolved 3D microvascular architecture comparable to two-photon microscopy while providing quantitative measurements of RBC flow and oxygenation. In live mice

SR-fPAM revealed redistribution of oxygen and hemodynamics across 3D microvascular networks following a single-vessel stroke. These findings establish SR-fPAM as an enabling tool that bridges a critical gap in oxygen-metabolism imaging and opens new avenues for studying microvascular health and disease with unprecedented functional insights.

关键词

Keywords

references

Andreone, B. J., Lacoste, B. & Gu, C. H. Neuronal and vascular interactions. Annu. Rev. Neurosci. 38 , 25–46 (2015)..

Hillman, E. M. C. Coupling mechanism and significance of the BOLD signal: a status report. Annu. Rev. Neurosci. 37 , 161–181 (2014)..

Grienberger, C. et al. Two-photon calcium imaging of neuronal activity. Nat. Rev. Methods Primers 2 , 67 (2022)..

Figley, C. R. & Stroman, P. W. The role(s) of astrocytes and astrocyte activity in neurometabolism, neurovascular coupling, and the production of functional neuroimaging signals. Eur. J. Neurosci. 33 , 577–588 (2011)..

Girouard, H. & Iadecola, C. Neurovascular coupling in the normal brain and in hypertension, stroke, and Alzheimer disease. J. Appl. Physiol. 100 , 328–335 (2006)..

Zhang, Z. K. et al. Imaging volumetric dynamics at high speed in mouse and zebrafish brain with confocal light field microscopy. Nat. Biotechnol. 39 , 74–83 (2021)..

Zhou, Q. Y. et al. Three-dimensional wide-field fluorescence microscopy for transcranial mapping of cortical microcirculation. Nat. Commun. 13 , 7969 (2022)..

Meng, G. H. et al. Ultrafast two-photon fluorescence imaging of cerebral blood circulation in the mouse brain in vivo. Proc. Natl. Acad. Sci. USA 119 , e2117346119 (2022)..

Fan, J.L. et al. High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics. Nat. Commun. 11 , 6020 (2020)..

Lecoq, J. et al. Simultaneous two-photon imaging of oxygen and blood flow in deep cerebral vessels. Nat. Med. 17 , 893–898 (2011)..

Yao, J. J. et al. High-speed label-free functional photoacoustic microscopy of mouse brain in action. Nat. Methods 12 , 407–410 (2015)..

Zhong, F. H. et al. High-speed wide-field multi-parametric photoacoustic microscopy. Opt. Lett. 45 , 2756–2759 (2020)..

Yao, J. J. & Wang, L. V. Photoacoustic microscopy. Laser Photonics Rev. 7 , 758–778 (2013)..

Jeon, S. et al. Review on practical photoacoustic microscopy. Photoacoustics 15 , 100141 (2019)..

Wang, L. V. & Hu, S. Photoacoustic tomography: in vivo imaging from organelles to organs. Science 335 , 1458–1462 (2012)..

Ma, J. et al. Grueneisen relaxation photoacoustic microscopy in vivo. J. Biomed. Opt. 21 , 066005 (2016)..

Yao, J. J. et al. Photoimprint photoacoustic microscopy for three-dimensional label-free subdiffraction imaging. Phys. Rev. Lett. 112 , 014302 (2014)..

Kim, J. et al. Super-resolution localization photoacoustic microscopy using intrinsic red blood cells as contrast absorbers. Light Sci. Appl. 8 , 103 (2019)..

Errico, C. et al. Ultrafast ultrasound localization microscopy for deep super-resolution vascular imaging. Nature 527 , 499–502 (2015)..

Renaudin, N. et al. Functional ultrasound localization microscopy reveals brain-wide neurovascular activity on a microscopic scale. Nat. Methods 19 , 1004–1012 (2022)..

Luís Dean-Ben, X. & Razansky, D. Localization optoacoustic tomography. Light Sci. Appl. 7 , 18004 (2018)..

Zhang, P. F. et al. In vivo superresolution photoacoustic computed tomography by localization of single dyed droplets. Light Sci. Appl. 8 , 36 (2019)..

Deán-Ben, X. L. et al. Deep optoacoustic localization microangiography of ischemic stroke in mice. Nat. Commun. 14 , 3584 (2023)..

Tzoumas, S. et al. Eigenspectra optoacoustic tomography achieves quantitative blood oxygenation imaging deep in tissues. Nat. Commun. 7 , 12121 (2016)..

Reeson, P., Choi, K. & Brown, C. E. VEGF signaling regulates the fate of obstructed capillaries in mous e cortex. eLife 7 , e33670 (2018)..

Li, H. et al. Disposable ultrasound-sensing chronic cranial windowby soft nanoimprinting lithography. Nat. Commun. 10 , 4277 (2019)..

Cao, R. et al. Functional and oxygen-metabolic photoacoustic microscopy of the awake mouse brain. NeuroImage 150 , 77–87 (2017)..

Wang, T. X. et al. Multiparametric photoacoustic microscopy of the mouse brain with 300-kHz A-line rate. Neurophotonics 3 , 045006 (2016)..

Nishimura, N. et al. Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke. Nat. Methods 3 , 99–108 (2006)..

Schaffer, C. B. et al. Two-photon imaging of cortical surface microvessels reveals a robust redistribution in blood flow after vascular occlusion. PLoS Biol. 4 , e22 (2006)..

Sun, N. D. et al. In vivo imaging of hemodynamic redistribution and arteriogenesis across microvascular network. Microcirculation 27 , e12598 (2020)..

Descloux, A., Grußmayer, K. S. & Radenovic, A. Parameter-free image resolution estimation based on decorrelation analysis. Nat. Methods 16 , 918–924 (2019)..

Wang, Y. M. et al. High-speed multi-parametric photoacoustic microscopy of cerebral hemodynamic and metabolic responses to acute hemodilution. Opt. Lett. 47 , 1988–1991 (2022)..

Guo, H. et al. Detachable head-mounted photoacoustic microscope in freely moving mice. Opt. Lett. 46 , 6055–6058 (2021)..

Kim, J. Y. et al. High-speed and high-SNR photoacoustic microscopy based on a galvanometer mirror in non-conducting liquid. Sci. Rep. 6 , 34803 (2016)..

Yao, J. J. et al. Wide-field fast-scanning photoacoustic microscopy based on a water-immersible MEMS scanning mirror. J. Biomed. Opt. 17 , 080505 (2012)..

Lan, B. X. et al. High-speed widefield photoacoustic microscopy of small-animal hemodynamics. Biomed. Opt. Express 9 , 4689–4701 (2018)..

Rust, M. J., Bates, M. & Zhuang, X. W. Sub-diffr action-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat. Methods 3 , 793–796 (2006)..

Couture, O. et al. Ultrasound localization microscopy and super-resolution: a state of the art. IEEE Trans. Ultrason. Ferroelectr. Freq. Control 65 , 1304–1320 (2018)..

Christensen-Jeffries, K. et al. In vivo acoustic super-resolution and super-resolved velocity mapping using microbubbles. IEEE Trans. Med. Imaging 34 , 433–440 (2015)..

Yang, X. et al. Photoacoustic imaging for monitoring of stroke diseases: a review. Photoacoustics 23 , 100287 (2021)..

Cao, R. et al. Photoacoustic microscopy of obesity-induced cerebrovascular alterations. NeuroImage 188 , 369–379 (2019)..

Gong, X. R. et al. Photoacoustic microscopy visualizes glioma-induced disruptions of cortical microvascular structure and function. J. Neural Eng. 19 , 026027 (2022)..

Guo, T. et al. Homogeneous-resolution photoacoustic microscopy for ultrawide field-of-view neurovascular imaging in Alzheimer's disease. Photoacoustics 31 , 100516 (2023)..

Zhu, W. M. et al. Neurovascular coupling mechanisms in health and neurovascular uncoupling in Alzheimer's disease. Brain 145 , 2276–2292 (2022)..

Ovsepian, S. V. et al. Pushing the boundaries of neuroimaging with optoacoustics. Neuron 96 , 966–988 (2017)..

Li, B. Q. et al. More homogeneous capillary flow and oxygenation in deeper cortical layers correlate with increased oxygen extraction. eLife 8 , e42299 (2019)..

Şencan, İ et al. Optical measurement of microvascular oxygenation and blood flow responses in awake mouse cortex during functional activation. J. Cereb. Blood Flow. Metab. 42 , 510–525 (2022)..

Ouyang, W. et al. Deep learning massively accelerates super-resolution localization microscopy. Nat. Biotechnol. 36 , 460–468 (2018)..

Liu, X. et al. Deep learning for ultrasound localization microscopy. IEEE Trans. Med. Imaging 39 , 3064–3078 (2020)..

Kim, J. et al. Deep learning acceleration of multiscale superresolution localization photoacoustic imaging. Light Sci. Appl. 11 , 131 (2022)..

Du, J. Y. et al. Recent progress in near-infrared photoacoustic imaging. Biosens. Bioelectron. 191 , 113478 (2021)..

Cao, R. et al. Optical-resolution photoacoustic microscopy with a needle-shaped beam. Nat. Photonics 17 , 89–95 (2023)..

Illingworth, J. & Kittler, J. A survey of the Hough transform. Comput. Vis. Graph. Image Process. 44 , 87–116 (1988)..

Views

Downloads

CSCD

Alert me when the article has been cited

Submit

Tools

Publicity Resources

Vectorial lasing with designable topological charges based on Möbius-like correspondence in quasi-BICs

Editors at the frontier: exploring advanced optical engineering in the UK

Light people: Professor Mengkun Liu

Dual-comb absolute ranging reaches orbit

Tunable structured laser over full spatial spectrum

Related Author

No data

Related Institution

No data

⁰