AutoStepfinder: A fast and automated step detection method for single-molecule analysis

Luuk Loeff, Jacob W.J. Kerssemakers, Chirlmin Joo, Cees Dekker

Research output: Contribution to journalArticlepeer-review

12 Scopus citations


Single-molecule techniques allow the visualization of the molecular dynamics of nucleic acids and proteins with high spatiotemporal resolution. Valuable kinetic information of biomolecules can be obtained when the discrete states within single-molecule time trajectories are determined. Here, we present a fast, automated, and bias-free step detection method, AutoStepfinder, that determines steps in large datasets without requiring prior knowledge on the noise contributions and location of steps. The analysis is based on a series of partition events that minimize the difference between the data and the fit. A dual-pass strategy determines the optimal fit and allows AutoStepfinder to detect steps of a wide variety of sizes. We demonstrate step detection for a broad variety of experimental traces. The user-friendly interface and the automated detection of AutoStepfinder provides a robust analysis procedure that enables anyone without programming knowledge to generate step fits and informative plots in less than an hour.

Original languageEnglish
Article number100256
Issue number5
StatePublished - 14 May 2021


  • AutoStepfinder
  • DSML 3: Development/Pre-production: Data science output has been rolled out/validated across multiple domains/problems
  • Stepfinder
  • biophysics
  • data analysis
  • fluorescence
  • magnetic tweezer
  • nanopore
  • optical tweezer
  • single molecule
  • step detection


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