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Aptamers as potential tools for super-resolution microscopy

Nature Methods volume 9, pages 938–939 (2012)Cite this article

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Epitopes must be labeled at high density for accurate super-resolution imaging, with the fluorophores as close as possible to their targets. Intuitively, it seems that small probes would be ideal for such efforts. This was demonstrated in a paper published recently in Nature Methods whose authors used 13-kDa camelid antibodies (nanobodies) to detect GFP-tagged proteins1. Super-resolution images obtained with nanobodies were far superior to those of conventional antibodies (∼150 kDa in size)1. However, the nanobodies used did not address endogenous, nontagged proteins. The GFP tag occasionally induces loss of function, multimerization or mislocalization. An additional issue that will specifically affect super-resolution imaging is the discontinuous labeling of structures containing both the GFP-tagged protein (seen by the GFP nanobodies) and the native copy (invisible to GFP nanobodies).

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Figure 1: Aptamers allow for better recognition of endosomes in super-resolution imaging.

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Acknowledgements

We thank K. Kröhnert for expert technical assistance, S. Wibbeke for assistance with experimental work and A. Schönle for the image acquisition software Imspector. The work was supported by a Starting Grant from the European Research Council, Program FP7 to S.O.R. (NANOMAP) and by funding provided by the US National Institutes of Health (5 R01 GM084703-02) and the Welch Foundation (F-1654) to A.D.E. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the US National Institutes of Health. T.W.G. acknowledges the support of the Interdisciplinary Center of Clinical Research at the University Hospital of the University of Erlangen-Nuremberg. M.L. acknowledges support from his Innovative Research Grant from Stand Up to Cancer, a program of the Entertainment Industry Foundation.

Author information

Authors and Affiliations

  1. STED Microscopy Group, European Neuroscience Institute, Deutsche Forschungsgemeinschaft Center for Molecular Physiology of the Brain/Excellence Cluster 171, Göttingen, Germany

    Felipe Opazo, Christina Schäfer & Silvio O Rizzoli

  2. Department of Neuro- and Sensory Physiology, University of Göttingen, Göttingen, Germany

    Felipe Opazo & Silvio O Rizzoli

  3. Department of Biochemistry, Albert Einstein College of Medicine, Bronx, New York, USA

    Matthew Levy

  4. Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas, USA

    Michelle Byrom

  5. Department of Optical Nanoscopy, Laser-Laboratory Göttingen e.V., Göttingen, Germany

    Claudia Geisler

  6. Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

    Claudia Geisler

  7. Department of Psychiatry and Psychotherapy, University of Erlangen, Erlangen, Germany

    Teja W Groemer

  8. Center for Systems and Synthetic Biology, Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas, USA

    Andrew D Ellington

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  1. Felipe Opazo
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Correspondence to Silvio O Rizzoli.

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Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–7 and Supplementary Methods (PDF 625 kb)

Supplementary Video 1

Live STED imaging of the c2 aptamer. Aptamer staining supplied sufficient fluorescence to be detected in both confocal or STED imaging, at a frequency of 2.8 frames per second. Note that the confocal and STED movies are not recorded in parallel. To avoid the bias introduced by bleaching, separate regions were imaged in STED or confocal mode. Scale bar, 500 nm. (MOV 8024 kb)

Supplementary Video 2

Live STED imaging of the A9 aptamer. STED and confocal movies were recorded exactly as for the c2 aptamer. Scale bar, 500 nm. (MOV 8724 kb)

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Opazo, F., Levy, M., Byrom, M. et al. Aptamers as potential tools for super-resolution microscopy. Nat Methods 9, 938–939 (2012). https://doi.org/10.1038/nmeth.2179

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