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Mid-infrared wide-field nanoscopy

Abstract

Mid-infrared (MIR) spectroscopy is widely recognized as a powerful, non-destructive method for chemical analysis. However, its utility is constrained by a micrometre-scale spatial resolution imposed by the long-wavelength MIR diffraction limit. This limitation has been recently overcome by MIR photothermal imaging, which detects photothermal effects induced in the vicinity of MIR absorbers using a visible-light microscope. Despite its promise, the full potential of its spatial resolving power has not been realized. Here we present an optimal implementation of wide-field MIR photothermal imaging to achieve high spatial resolution. This was accomplished by employing single-objective synthetic-aperture quantitative phase imaging with synchronized subnanosecond MIR and visible light sources, effectively suppressing the resolution-degradation effect caused by photothermal heat diffusion. We demonstrated far-field MIR spectroscopic imaging with a spatial resolution limited by the visible diffraction, down to 120 or 175 nm in terms of the Nyquist–Shannon sampling theorem or full-width at half-maximum of the point spread function, respectively, in the MIR region of 3.12–3.85 μm (2,600–3,200 cm−1). This technique—through the use of a shorter visible wavelength and/or a higher objective numerical aperture—holds the potential to achieve a spatial resolution of less than 100 nm, thus paving the way for MIR wide-field nanoscopy.

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Fig. 1: Single-objective synthetic-aperture MIP-QPI.
Fig. 2: Mid-infrared spectroscopic imaging with 175 nm FWHM or 120 nm Nyquist–Shannon sampling resolution.
Fig. 3: Dissecting bacterial groups by high-spatial-resolution MIR spectroscopic imaging.

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Data availability

An example set of data for SOSA-MIP-QPI can be found at https://github.com/takuroideguchi/SOSA-MIP-QPI_2024_demo. Other data presented in this work are available from the corresponding author on reasonable request.

Code availability

An example of the analysis code for SOSA-MIP-QPI can be found at https://github.com/takuroideguchi/SOSA-MIP-QPI_2024_demo. Other codes used in this work are available from the corresponding author on reasonable request.

References

  1. Pavlovetc, I. M., Aleshire, K., Hartland, G. V. & Kuno, M. Approaches to mid-infrared, super-resolution imaging and spectroscopy. Phys. Chem. Chem. Phys. 22, 4313–4325 (2020).

    Article  Google Scholar 

  2. Raghunathan, V., Han, Y., Korth, O., Ge, N. H. & Potma, E. O. Rapid vibrational imaging with sum frequency generation microscopy. Opt. Lett. 36, 3891–3893 (2011).

    Article  ADS  Google Scholar 

  3. Hanninen, A. M., Prince, R. C., Ramos, R., Plikus, M. V. & Potma, E. O. High-resolution infrared imaging of biological samples with third-order sum-frequency generation microscopy. Biomed. Opt. Express 9, 4807–4817 (2018).

    Article  Google Scholar 

  4. Shi, J. et al. High-resolution, high-contrast mid-infrared imaging of fresh biological samples with ultraviolet-localized photoacoustic microscopy. Nat. Photon. 13, 609–615 (2019).

    Article  ADS  Google Scholar 

  5. Bai, Y., Yin, J. & Cheng, J. X. Bond-selective imaging by optically sensing the MIR-infrared photothermal effect. Sci. Adv. 7, eabg1559 (2021).

    Article  ADS  Google Scholar 

  6. Cotte, Y. et al. Marker-free phase nanoscopy. Nat. Photon. 7, 113–117 (2013).

    Article  ADS  Google Scholar 

  7. Zhang, D. et al. Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution. Sci. Adv. 2, e1600521 (2016).

    Article  ADS  Google Scholar 

  8. Li, Z., Aleshire, K., Kuno, M. & Hartland, G. V. Super-resolution far-field infrared imaging by photothermal heterodyne imaging. J. Phys. Chem. B 121, 8838–8846 (2017).

    Article  Google Scholar 

  9. Samolis, P. D. & Sander, M. Y. Phase-sensitive lock-in detection for high-contrast mid-infrared photothermal imaging with sub-diffraction limited resolution. Opt. Express 27, 2643–2655 (2019).

    Article  ADS  Google Scholar 

  10. Lim, J. M. et al. Cytoplasmic protein imaging with mid-infrared photothermal microscopy: cellular dynamics of live neurons and oligodendrocytes. J. Phys. Chem. Lett. 10, 2857–2861 (2019).

    Article  Google Scholar 

  11. Toda, K., Tamamitsu, M., Nagashima, Y., Horisaki, R. & Ideguchi, T. Molecular contrast on phase-contrast microscope. Sci. Rep. 9, 9957 (2019).

    Article  ADS  Google Scholar 

  12. Li, X. et al. Fingerprinting a living cell by Raman integrated mid-infrared photothermal microscopy. Anal. Chem. 91, 10750–10756 (2019).

    Article  Google Scholar 

  13. Bauld, R., Choi, D. Y. W., Bazylewski, P., Divigalpitiya, R. & Fanchini, G. Thermo-optical characterization and thermal properties of graphene-polymer composites: a review. J. Mater. Chem. C 6, 2901–2914 (2018).

    Article  Google Scholar 

  14. Marín, E. The role of thermal properties in periodic time-varying phenomena. Eur. J. Phys. 28, 429–445 (2007).

    Article  ADS  Google Scholar 

  15. Fu, P. et al. Super-resolution imaging of non-fluorescent molecules by photothermal relaxation localization microscopy. Nat. Photon. 17, 330–337 (2023).

    Article  ADS  Google Scholar 

  16. Bai, Y. et al. Ultrafast chemical imaging by wide-field photothermal sensing of infrared absorption. Sci. Adv. 5, eaav7127 (2019).

    Article  ADS  Google Scholar 

  17. Tamamitsu, M., Toda, K., Horisaki, R. & Ideguchi, T. Quantitative phase imaging with molecular vibrational sensitivity. Opt. Lett. 44, 3729–3732 (2019).

    Article  ADS  Google Scholar 

  18. Zhang, D. et al. Bond-selective transient phase imaging via sensing of the infrared photothermal effect. Light Sci. Appl. 8, 116 (2019).

    Article  ADS  Google Scholar 

  19. Tamamitsu, M. et al. Label-free biochemical quantitative phase imaging with mid-infrared photothermal effect. Optica 7, 359–366 (2020).

    Article  ADS  Google Scholar 

  20. Toda, K., Tamamitsu, M. & Ideguchi, T. Adaptive dynamic range shift (ADRIFT) quantitative phase imaging. Light Sci. Appl. 10, 1 (2021).

    Article  ADS  Google Scholar 

  21. Yurdakul, C., Zong, H., Bai, Y., Cheng, J. X. & Ünlü, M. S. Bond-selective interferometric scattering microscopy. J. Phys. D 54, 364002 (2021).

    Article  ADS  Google Scholar 

  22. Ishigane, G. et al. Mid-infrared photothermal single-live-cell imaging beyond video rate. Light Sci. Appl. 12, 174 (2023).

    Article  ADS  Google Scholar 

  23. Zharov, V. P. & Lapotko, D. O. Photothermal imaging of nanoparticles and cells. IEEE J. Sel. Top. Quantum Electron. 11, 733–751 (2005).

    Article  ADS  Google Scholar 

  24. Jaumot, J., Gargallo, R., Juan, A. & Tauler, R. A graphical user-friendly interface for MCR-ALS: a new tool for multivariate curve resolution in Matlab. Chemom. Intell. Lab. Syst. 76, 101–110 (2005).

    Article  Google Scholar 

  25. Zhang, C. et al. Bacterial lipid droplets bind to DNA via an intermediary protein that enhances survival under stress. Nat. Commun. 8, 15979 (2017).

    Article  ADS  Google Scholar 

  26. Alvarez, H. M., Silva, R. A., Herrero, M., Hernández, M. A. & Villalba, M. S. Metabolism of triacylglycerols in Rhodococcus species: insights from physiology and molecular genetics. J. Mol. Biochem. 2, 67–78 (2012).

    Google Scholar 

  27. Paviva, E. M. & Schmidt, F. M. Ultrafast widefield mid-infrared photothermal heterodyne imaging. Anal. Chem. 94, 14242–14250 (2022).

    Article  Google Scholar 

  28. Yin, J. et al. Video-rate mid-infrared photothermal imaging by single-pulse photothermal detection per pixel. Sci. Adv. 9, eadg8814 (2023).

    Article  Google Scholar 

  29. Baek, Y. S., Hugonnet, H. & Park, Y. K. Pupil-aberration calibration with controlled illumination for quantitative phase imaging. Opt. Express 29, 22127–22135 (2021).

    Article  ADS  Google Scholar 

  30. Guo, Z., Bai, Y., Zhang, M., Lan, L. & Cheng, J. X. High-throughput antimicrobial susceptibility testing of Escherichi coli by wide-field mid-infrared photothermal imaging of protein synthesis. Anal. Chem. 95, 2238–2244 (2023).

    Article  Google Scholar 

  31. Gao, P. & Yuan, C. Resolution enhancement of digital holographic microscopy via synthetic aperture: a review. Light: Adv. Manuf. 3, 1–16 (2022).

    Google Scholar 

  32. Andika, M., Chen, G. C. K. & Vasudevan, S. Excitation temporal pulse shape and probe beam size effect on pulsed photothermal lens of single particle. J. Opt. Soc. Am. B 27, 796–805 (2010).

    Article  ADS  Google Scholar 

  33. Smith, G.D. Numerical Solution of Partial Differential Equations: Finite Difference Methods (Oxford Univ. Press, 1985).

  34. Blum O. & Shaked, N.T. Prediction of photothermal phase signatures from arbitrary plasmonic nanoparticles and experimental verification. Light Sci. Appl. 4, e322 (2015).

  35. Water—Thermal Diffusivity (EngineeringToolBox, accessed 27 September 2023); https://www.engineeringtoolbox.com/water-steam-thermal-diffusivity-d_2058.html

  36. Goodman, J.W. Introduction to Fourier Optics (Roberts and Company Publishers, 2005).

  37. Coleman, N. V., Mattes, T. E., Gosset, J. M. & Spain, J. C. Biodegradation of cis-dichloroethene as the sole carbon source by a β-proteobacterium. Appl. Environ. Microbiol. 68, 2726–2730 (2002).

    Article  ADS  Google Scholar 

  38. GraphPad Prism v.9.5.1 for Mac. (GraphPad Software, 2024).

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Acknowledgements

We thank G. Ishigane for fruitful discussions. This work was financially supported by Japan Society for the Promotion of Science (grant nos. 20H00125 and 23H00273 to T.I.), JST PRESTO (grant no. JPMJPR17G2 to T.I.), Precise Measurement Technology Promotion Foundation (to T.I.), Research Foundation for Opto-Science and Technology (to T.I.), Nakatani Foundation (granted to T.I.) and a UTEC-Utokyo FSI Research Grant (to T.I.). Fabrication of the custom-made resolution test chart was performed using the apparatus at the Takeda Clean Room of d.lab at The University of Tokyo.

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Authors and Affiliations

Authors

Contributions

M.T. conceived the concept of SOSA-MIP-QPI and designed the system. M.T. and K.T. performed thermal conduction calculations. M.T. performed the imaging-related simulations. V.R.B. constructed the light sources. H.S. and M.F. wrote the automated data acquisition program. K.K. fabricated the nanoscale spatial-resolution test chart. S.O. provided the E. coli cells. M.T. prepared the R. jostii RHA1 cells. M.T., K.T. and M.F. performed the experiments. M.T. analysed the data. T.I. supervised the work. M.T., K.T. and T.I. wrote the manuscript.

Corresponding author

Correspondence to Takuro Ideguchi.

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Competing interests

M.T., K.T. and T.I. are inventors of patents related to MIP-QPI. The remaining authors declare no competing interests.

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Nature Photonics thanks the anonymous reviewers for their contribution to the peer review of this work.

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Supplementary Notes 1–13 and Figs 1–13.

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Tamamitsu, M., Toda, K., Fukushima, M. et al. Mid-infrared wide-field nanoscopy. Nat. Photon. (2024). https://doi.org/10.1038/s41566-024-01423-0

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