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Analysing errors in single-molecule localisation microscopy

Research output: Contribution to journalShort surveypeer-review

Original languageEnglish
Article number105931
JournalInternational Journal of Biochemistry and Cell Biology
PublishedMay 2021

Bibliographical note

Funding Information: This work was supported by the BBSRC (BB/R021767/1) and the MRC (MR/R008264/1). IC acknowledges support from an EPSRC studentship. Publisher Copyright: © 2021 Elsevier Ltd Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

King's Authors


In single molecule localisation microscopy (SMLM) a super-resolution image of the distribution of fluorophores in the sample is built up from the localised positions of many individual molecules. It has become widely used due to its experimental simplicity and the high resolution that can be achieved. However, the factors which limit resolution in a reconstructed image, and the artefacts which can be present, are completely different to those present in standard fluorescent microscopy techniques. Artefacts may be difficult for users to identify, particularly as they can cause images to appear falsely sharp, an effect called artificial sharpening. Here we discuss the different sources of error and bias in SMLM, and the methods available for avoiding or detecting them.

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