Abstract
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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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.
References
Boyer, D., Tamarat, P., Maali, A., Lounis, B. & Orrit, M. Photothermal imaging of nanometer-sized metal particles among scatterers. Science 297, 1160–1163 (2002).
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).
Zhao, M., Wang, X. F. & Nolte, D. D. Molecular interferometric imaging. Opt. Express 16, 7102–7118 (2008).
Arbouet, A. et al. Direct measurement of the single-metal-cluster optical absorption. Phys. Rev. Lett. 93, 127401 (2004).
Min, W. et al. Imaging chromophores with undetectable fluorescence by stimulated emission microscopy. Nature 461, 1105–1109 (2009).
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).
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).
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).
Zumofen, G., Mojarad, N. M., Sandoghdar, V. & Agio, M. Perfect reflection of light by an oscillating dipole. Phys. Rev. Lett. 101, 180404 (2008).
Kohl, C., Becker, S. & Müllen, K. Bis(rylenedicarboximide)-a,d-1,5-diaminoanthraquinones as unique infrared absorbing dyes. Chem. Commun. 2778–2779 (2002).
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).
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).
Kukura, P., Celebrano, M., Renn, A. & Sandoghdar, V. Imaging a single quantum dot when it is dark. Nano Lett. 9, 926–929 (2009).
Brinks, D. et al. Visualizing and controlling vibrational wave packets of single molecules. Nature 465, 905–908 (2010).
Giessibl, F. J. Advances in atomic force microscopy. Rev. Mod. Phys. 75, 949–983 (2003).
Sugimoto, Y. et al. Chemical identification of individual surface atoms by atomic force microscopy. Nature 446, 64–67 (2007).
Plakhotnik, T. & Palm, V. Interferometric signatures of single molecules. Phys. Rev. Lett. 87, 183602 (2001).
Gerhardt, I. et al. Strong extinction of a laser beam by a single molecule. Phys. Rev. Lett. 98, 033601 (2007).
Kukura, P. et al. High-speed nanoscopic tracking of the position and orientation of a single virus. Nat. Methods 6, 923–927 (2009).
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).
Sannomiya, T., Hafner, C. & Voros, J. In situ sensing of single binding events by localized surface plasmon resonance. Nano Lett. 8, 3450–3455 (2008).
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).
Acknowledgements
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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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.
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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). https://doi.org/10.1038/nphoton.2010.290
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DOI: https://doi.org/10.1038/nphoton.2010.290
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