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Single-molecule imaging by optical absorption

An Author Correction to this article was published on 12 April 2018

This article has been updated


To date, optical studies of single molecules at room temperature have relied on the use of materials with high fluorescence quantum yield combined with efficient spectral rejection of background light. To extend single-molecule studies to a much larger pallet of substances that absorb but do not fluoresce, scientists have explored the photothermal effect1, interferometry2,3, direct attenuation4 and stimulated emission5. Indeed, very recently, three groups have succeeded in achieving single-molecule sensitivity in absorption6,7,8. Here, we apply modulation-free transmission measurements known from absorption spectrometers to image single molecules under ambient conditions both in the emissive and strongly quenched states. We arrive at quantitative values for the absorption cross-section of single molecules at different wavelengths and thereby set the ground for single-molecule absorption spectroscopy. Our work has important implications for research ranging from absorption and infrared spectroscopy to sensing of unlabelled proteins at the single-molecule level.

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Figure 1: Transmission microscopy for single-molecule imaging.
Figure 2: Single-molecule absorption imaging and characterization.
Figure 3: Background rejection by polarization rotation.
Figure 4: Dual-colour single-molecule spectroscopy.
Figure 5: Imaging of non-fluorescent molecules.

Change history

  • 12 April 2018

    In the Supplementary Video initially published with this Letter, the right-hand panel displaying the fluorescence emission was not showing on some video players due to a formatting problem; this has now been fixed. The video has also now been amended to include colour scale bars for both the left- (differential transmission signal) and right-hand panels.


  1. Boyer, D., Tamarat, P., Maali, A., Lounis, B. & Orrit, M. Photothermal imaging of nanometer-sized metal particles among scatterers. Science 297, 1160–1163 (2002).

    ADS  Article  Google Scholar 

  2. Hwang, J., Fejer, M. M. & Moerner, W. E. Scanning interferometric microscopy for the detection of ultrasmall phase shifts in condensed matter. Phys. Rev. A 73, 021802 (2006).

    ADS  Article  Google Scholar 

  3. Zhao, M., Wang, X. F. & Nolte, D. D. Molecular interferometric imaging. Opt. Express 16, 7102–7118 (2008).

    ADS  Article  Google Scholar 

  4. Arbouet, A. et al. Direct measurement of the single-metal-cluster optical absorption. Phys. Rev. Lett. 93, 127401 (2004).

    ADS  Article  Google Scholar 

  5. Min, W. et al. Imaging chromophores with undetectable fluorescence by stimulated emission microscopy. Nature 461, 1105–1109 (2009).

    ADS  Article  Google Scholar 

  6. Kukura, P., Celebrano, M., Renn, A. & Sandoghdar, V. Single-molecule sensitivity in optical absorption at room temperature. J. Phys. Chem. Lett. 1, 3323–3327 (2010).

    Article  Google Scholar 

  7. Gaiduk, A., Yorulmaz, M., Ruijgrok, P. V. & Orrit, M. Room-temperature detection of a single molecule's absorption by photothermal contrast. Science 330, 353–356 (2010).

    ADS  Article  Google Scholar 

  8. Chong, S., Min, W. & Xie, S. Ground-state depletion microscopy: detection sensitivity of single-molecule optical absorption at room temperature. J. Phys. Chem. Lett. 1, 3316–3322 (2010).

    Article  Google Scholar 

  9. Zumofen, G., Mojarad, N. M., Sandoghdar, V. & Agio, M. Perfect reflection of light by an oscillating dipole. Phys. Rev. Lett. 101, 180404 (2008).

    ADS  Article  Google Scholar 

  10. Kohl, C., Becker, S. & Müllen, K. Bis(rylenedicarboximide)-a,d-1,5-diaminoanthraquinones as unique infrared absorbing dyes. Chem. Commun. 2778–2779 (2002).

  11. Mais, S. et al. Terrylenediimide: a novel fluorophore for single-molecule spectroscopy and microscopy from 1.4 K to room temperature. J. Phys. Chem. A 101, 8435–8440 (1997).

    Article  Google Scholar 

  12. Jacobsen, V., Stoller, P., Brunner, C., Vogel, V. & Sandoghdar, V. Interferometric optical detection and tracking of very small gold nanoparticles at a water–glass interface. Opt. Express 14, 405–414 (2006).

    ADS  Article  Google Scholar 

  13. Kukura, P., Celebrano, M., Renn, A. & Sandoghdar, V. Imaging a single quantum dot when it is dark. Nano Lett. 9, 926–929 (2009).

    ADS  Article  Google Scholar 

  14. Brinks, D. et al. Visualizing and controlling vibrational wave packets of single molecules. Nature 465, 905–908 (2010).

    ADS  Article  Google Scholar 

  15. Giessibl, F. J. Advances in atomic force microscopy. Rev. Mod. Phys. 75, 949–983 (2003).

    ADS  Article  Google Scholar 

  16. Sugimoto, Y. et al. Chemical identification of individual surface atoms by atomic force microscopy. Nature 446, 64–67 (2007).

    ADS  Article  Google Scholar 

  17. Plakhotnik, T. & Palm, V. Interferometric signatures of single molecules. Phys. Rev. Lett. 87, 183602 (2001).

    ADS  Article  Google Scholar 

  18. Gerhardt, I. et al. Strong extinction of a laser beam by a single molecule. Phys. Rev. Lett. 98, 033601 (2007).

    ADS  Article  Google Scholar 

  19. Kukura, P. et al. High-speed nanoscopic tracking of the position and orientation of a single virus. Nat. Methods 6, 923–927 (2009).

    Article  Google Scholar 

  20. Armani, A. M., Kulkarni, R. P., Fraser, S. E., Flagan, R. C. & Vahala, K. J. Label-free, single-molecule detection with optical microcavities. Science 317, 783–787 (2007).

    ADS  Article  Google Scholar 

  21. Sannomiya, T., Hafner, C. & Voros, J. In situ sensing of single binding events by localized surface plasmon resonance. Nano Lett. 8, 3450–3455 (2008).

    ADS  Article  Google Scholar 

  22. Wrigge, G., Hwang, J., Gerhardt, I., Zumofen, G. & Sandoghdar, V. Exploring the limits of single emitter detection in fluorescence and extinction. Opt. Express 16, 17358–17365 (2008).

    ADS  Article  Google Scholar 

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The authors thank G. Grassi for synthesis of TDI. This work was supported by ETH Zurich and the Swiss National Foundation.

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Authors and Affiliations



M.C. performed the experiments. M.C. and P.K. analysed the data. A.R. and V.S. supervised the project. V.S., P.K. and M.C. wrote the manuscript.

Corresponding author

Correspondence to Vahid Sandoghdar.

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The authors declare no competing financial interests.

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Celebrano, M., Kukura, P., Renn, A. et al. Single-molecule imaging by optical absorption. Nature Photon 5, 95–98 (2011).

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