Microscopy and its focal switch

Abstract

Until not very long ago, it was widely accepted that lens-based (far-field) optical microscopes cannot visualize details much finer than about half the wavelength of light. The advent of viable physical concepts for overcoming the limiting role of diffraction in the early 1990s set off a quest that has led to readily applicable and widely accessible fluorescence microscopes with nanoscale spatial resolution. Here I discuss the principles of these methods together with their differences in implementation and operation. Finally, I outline potential developments.

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Figure 1: Memorial erected in Ernst Abbe's honor displaying his equation describing the diffraction resolution limit, located in front of the physiology building of the University of Jena, Germany.
Figure 2: Fluorescence switching strategies for super-resolution image assembly.

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Acknowledgements

I thank E. Rittweger and B. Rankin for help in preparing the figures and various suggestions for improving the presentation. Critical reading by A. Schönle, and also by S. Jakobs, C. Eggeling and J. Jethwa, is gratefully acknowledged.

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Correspondence to Stefan W Hell.

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Hell, S. Microscopy and its focal switch. Nat Methods 6, 24–32 (2009). https://doi.org/10.1038/nmeth.1291

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