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Single-particle analysis for fluorescence nanoscopy

Nature Methods volume 15, pages 771–772 (2018)Cite this article

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Building on methods from electron microscopy, researchers are applying nanoscale fluorescence imaging to address questions in structural biology.

Advanced fluorescence microscopy methods achieve nanometer-scale resolution, which makes them capable, in principle, of probing the structure and composition of macromolecular complexes1. However, despite their high resolution, fluorescence nanoscopy images often appear incomplete, owing to partial labeling of the sample, or are noisy as a result of the limited brightness and stability of the fluorophores. These factors can make the measurement of structural parameters highly challenging. Is there a way to boost image quality further, without compromising resolution? Fortunately the answer is yes, and the solution lies in the power of image averaging, embodied by a suite of methods collectively known as single-particle analysis (SPA)2,3. These methods have already begun to find applications in fluorescence nanoscopy, particularly in the analysis of structures with high symmetry4,5,6. In this issue of Nature Methods, two new studies7,8 advance this approach further, bringing sophisticated SPA techniques to bear on biological structure determination by super-resolution fluorescence microscopy.

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Fig. 1: Single-particle analysis of multicolor SMLM images.

Kim Caesar/Springer Nature, based on images in ref. 8.

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  1. Department of Nanobiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany

    Mark Bates

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  1. Mark Bates
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Correspondence to Mark Bates.

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Bates, M. Single-particle analysis for fluorescence nanoscopy. Nat Methods 15, 771–772 (2018). https://doi.org/10.1038/s41592-018-0151-7

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