Super-Resolution Imaging

Abstract

Imaging using light microscopy is generally limited in resolution to approximately 250 nm. To overcome that limit, and see more detail, several different techniques have now been developed. This chapter introduces these techniques and explores how they can be used in live-cell imaging. After defining resolution and the resolution limit of a conventional widefield or confocal microscope, we begin by reviewing techniques that double the potential resolving power of such microscopes, including structured illumination microscopy (SIM), and pixel reassignment approaches such as Airyscan and instant SIM. We describe how STED (stimulated emission depletion) microscopy, which improves resolution by fivefold or more, is now being adopted successfully for live-cell imaging. We also discuss the use of single-molecule localisation microscopy techniques, which can surpass the resolution limit substantially, in live-cell imaging, including for tracking single molecules and in combination with light-sheet microscopy. Finally, we discuss how computational approaches can be used to improve spatial and temporal resolution across these techniques.

Metadata

Item Type:	Book Section
Authors/Creators:	Curd, A. Rahmani, A. Umney, O. Ponjavic, A. Peckham, M. https://orcid.org/0000-0002-3754-2028
Editors:	Linder, S. Wells, C.M.
Copyright, Publisher and Additional Information:	© 2026 John Wiley & Sons Ltd. This is the peer reviewed version of the following chapter: Curd, A., Rahmani, A., Umney, O., Ponjavic, A. and Peckham, M. (2026). Super-Resolution Imaging . In Live Biological Imaging Across Scales (eds S. Linder and C.M. Wells), which has been published in final form at https://doi.org/10.1002/9781394232413.ch1. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.
Keywords:	super-resolution structured illumination microscopy reassignment STED single-molecule localisation single-molecule tracking light-sheet microscopy fluorescence, dyes, image analysis
Dates:	Accepted: 29 January 2025 Published (online): 27 March 2026 Published: 2 June 2026
Institution:	The University of Leeds
Academic Units:	The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Cell Biology (Leeds)
Date Deposited:	18 Feb 2025 09:36
Last Modified:	20 Apr 2026 16:45
Status:	Published
Publisher:	Wiley
Identification Number:	10.1002/9781394232413.ch1
Open Archives Initiative ID (OAI ID):	oai:eprints.whiterose.ac.uk:222745

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Super-Resolution Imaging

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