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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.


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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.

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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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