Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells

Tatsuya Osaki; W. David Lee; Xiang Zhang; Rebecca E. Zubajlo; Mercedes Balcells-Camps; Elazer R. Edelman; Brian W. Anthony; Mriganka Sur; Peter T. C. So

doi:10.1038/s41377-025-01895-x

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Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells

Original Articles

- Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells
- Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells
- LSA 2025年14卷第9期页码：2784-2798
- Affiliations：
  
  1.Picower Institute for Learning and Memory, Massachusetts Institute of Technology, Cambridge, MA, USA
  2.Whitehead Institute for Biomedical Research, Massachusetts Institute of Technology, Cambridge, MA, USA
  3.Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
  4.Institute of Medical Engineering and Science, Massachusetts Institute of Technology, Cambridge, MA, USA
  5.Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA
- Author bio：
  
  Brian W. Anthony (banthony@mit.edu)
  Mriganka Sur (msur@mit.edu)
  Peter T. C. So (ptso@mit.edu)
- Funds：
- DOI：10.1038/s41377-025-01895-x
  中图分类号：
- 收稿日期：2024-12-24，
  
  修回日期：2025-03-21，
  
  录用日期：2025-05-16，
  
  网络出版日期：2025-08-07，
  
  纸质出版日期：2025-09-30
- Accepted：
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Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells[J]. LSA, 2025,14(9):2784-2798.

Osaki, T. et al. Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells. Light: Science & Applications, 14, 2784-2798 (2025).
Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells[J]. LSA, 2025,14(9):2784-2798. DOI： 10.1038/s41377-025-01895-x.

Osaki, T. et al. Multi-photon, label-free photoacoustic and optical imaging of NADH in brain cells. Light: Science & Applications, 14, 2784-2798 (2025). DOI： 10.1038/s41377-025-01895-x.

摘要

Abstract

Label-free detection of biological events at single-cell resolution in the brain can non-invasively capture brain status for medical diagnosis and basic neuroscience research. NADH is an universal coenzyme that not only plays a central role in cellular metabolism but may also be used as a biomarker to capture metabolic processes in brain cells and structures. We have developed a new label-free

multiphoton photoacoustic microscope (LF-MP-PAM) with a near-infrared femtosecond laser to observe endogenous NAD(P)H in living cells. The imaging depth of NAD(P)H in tissues with all-optical methods is limited to ~100 μm in brain tissue by the strong absorption of the near-ultraviolet fluorescence. Here

acoustic detection of the thermal signature of multi-photon (three-photon) excitation of NAD(P)H

a low quantum yield fluorophore

allows detection at an unprecedented depth while the focused excitation ensures high spatial resolution. We validated the photoacoustic detection of NAD(P)H by monitoring an increase in intracellular NAD(P)H in HEK293T cells and HepG2 cells incubated in NADH solution. We also demonstrated the detection of endogenous NAD(P)H photoacoustic signals in brain slices to 700 μm depth and in cerebral organoids to 1100 μm depth. Finally

we developed and demonstrated simultaneous photoacoustic and optical imaging of NAD(P)H in brain cells with a real-time image acquisition and processing pipeline. This approach could open a new door to monitor brain metabolic changes during development and disease

and changes due to neuronal activity

at single-cell level deep in the brains of both humans and animals.

关键词

Keywords

references

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