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Reply to: Assessment of 3D MINFLUX data for quantitative structural biology in cells

Nature Methods volume 20, pages 52–54 (2023)Cite this article

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The Original Article was published on 15 December 2022

replying to: Prakash, K. & Curd, A. P. Nat. Methods https://doi.org/10.1038/s41592-022-01694-x (2022).

Prakash and Curd provide a re-analysis1 of individual datasets taken from our report2 demonstrating MINFLUX three-dimensional (3D) imaging in cells. Their evaluation confirms the unique MINFLUX2,3 localization precision of σ = 1–3 nm standard deviation. We appreciate their confirmation and also welcome the opportunity to clarify their additional points.

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Fig. 1: Prakash and Curd neglected the tilts of individual NPCs in non-flat nuclear membranes.

Data availability

Source data are provided with this paper.

Code availability

The analysis code is available in Supplementary Software.

References

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Acknowledgements

We thank S. Schnorrenberg for providing the MINFLUX data of Supplementary Fig. 1, taken during a recent EMBL course at the EMBL Imaging Centre, Heidelberg.

Author information

Authors and Affiliations

  1. Department of NanoBiophotonics, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany

    Klaus C. Gwosch, Francisco Balzarotti, Jasmin K. Pape, Steffen J. Sahl & Stefan W. Hell

  2. Research Institute of Molecular Pathology, Vienna, Austria

    Francisco Balzarotti

  3. Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany

    Philipp Hoess, Jan Ellenberg & Jonas Ries

  4. Abberior Instruments GmbH, Göttingen, Germany

    Ulf Matti & Roman Schmidt

  5. Department of Optical Nanoscopy, Max Planck Institute for Medical Research, Heidelberg, Germany

    Stefan W. Hell

Authors
  1. Klaus C. Gwosch
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  2. Francisco Balzarotti
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  3. Jasmin K. Pape
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  4. Philipp Hoess
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  5. Jan Ellenberg
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  6. Jonas Ries
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  7. Ulf Matti
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  8. Roman Schmidt
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  9. Steffen J. Sahl
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  10. Stefan W. Hell
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Contributions

The reply was written by K.C.G., F.B., J.K.P., P.H., J.E., J.R., U.M., R.S., S.J.S. and S.W.H. F.B. and S.J.S. performed data analysis. U.M. and R.S. recorded MINFLUX data.

Corresponding author

Correspondence to Stefan W. Hell.

Ethics declarations

Competing interests

S.W.H. is a co-founder of the company Abberior Instruments, which commercializes super-resolution microscopy systems, including MINFLUX. The Max Planck Society holds patents on the principles, embodiments and procedures of MINFLUX, benefiting S.W.H., K.C.G. and F.B. U.M. and R.S. are employees of Abberior Instruments GmbH.

Peer review

Peer review information

Nature Methods thanks Bernd Rieger, and the other, anonymous, reviewers for their contribution to the peer review of this work. Primary Handling editor: Rita Strack, in collaboration with the Nature Methods team.

Supplementary information

Supplementary Information

Supplementary Fig. 1.

Reporting Summary

Supplementary Software

Particle fitting of pre-clustered NPCs in MINFLUX data in Gwosch et al. (2020).

Source data

Source Data Fig. 1c

Localizations in units of nanometer.

Source Data Fig. 1e

Localizations in units of nanometer.

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Gwosch, K.C., Balzarotti, F., Pape, J.K. et al. Reply to: Assessment of 3D MINFLUX data for quantitative structural biology in cells. Nat Methods 20, 52–54 (2023). https://doi.org/10.1038/s41592-022-01695-w

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