Quantitative fibre analysis of single-molecule localization microscopy data

Research output: Contribution to journalArticlepeer-review


  • Ruby Peters
  • Juliette Griffié
  • Garth L. Burn
  • David J. Williamson
  • Dylan Owen

Colleges, School and Institutes

External organisations

  • Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College, London
  • Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany


Single molecule localization microscopy (SMLM) methods produce data in the form of a spatial point pattern (SPP) of all localized emitters. Whilst numerous tools exist to quantify molecular clustering in SPP data, the analysis of fibrous structures has remained understudied. Taking the SMLM localization coordinates as input, we present an algorithm capable of tracing fibrous structures in data generated by SMLM. Based upon a density parameter tracing routine, the algorithm outputs several fibre descriptors, such as number of fibres, length of fibres, area of enclosed regions and locations and angles of fibre branch points. The method is validated in a variety of simulated conditions and experimental data acquired using the image reconstruction by integrating exchangeable single-molecule localization (IRIS) technique. For this, the nanoscale architecture of F-actin at the T cell immunological synapse in both untreated and pharmacologically treated cells, designed to perturb actin structure, was analysed.


Original languageEnglish
Article number10418
JournalScientific Reports
Issue number1
Publication statusPublished - 10 Jul 2018