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Related Experiment Video

Updated: Nov 16, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
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Analysing errors in single-molecule localisation microscopy.

Ishan Costello1, Susan Cox1

  • 1Randall Centre for Cell & Molecular Biophysics, Guy's Campus, King's College London, UK.

The International Journal of Biochemistry & Cell Biology
|February 20, 2021
PubMed
Summary
This summary is machine-generated.

Single molecule localisation microscopy (SMLM) achieves super-resolution by localising individual fluorophores. This review discusses SMLM

Keywords:
Data assessmentSingle-molecule localisation microscopySuper-resolution

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Area of Science:

  • Microscopy
  • Biophysics
  • Super-resolution imaging

Background:

  • Single Molecule Localisation Microscopy (SMLM) is a powerful super-resolution technique.
  • It reconstructs images from the positions of individual fluorescent molecules.
  • SMLM offers experimental simplicity and high resolution.

Purpose of the Study:

  • To discuss factors limiting resolution in SMLM.
  • To identify and explain artefacts unique to SMLM.
  • To present methods for avoiding or detecting SMLM errors and biases.

Main Methods:

  • Review of SMLM principles and limitations.
  • Analysis of common SMLM artefacts, including artificial sharpening.
  • Discussion of error sources and bias in SMLM data.

Main Results:

  • SMLM resolution limitations and artefacts differ from standard fluorescence microscopy.
  • Artefacts can be subtle and may cause artificial sharpening, leading to misinterpretation.
  • Various sources of error and bias can affect SMLM image quality.

Conclusions:

  • Understanding SMLM-specific artefacts is crucial for accurate image interpretation.
  • Methods for artefact detection and error avoidance are essential for reliable SMLM results.
  • This review provides guidance for users to identify and mitigate SMLM limitations.