Single-Molecule FRET X

Mike Filius, Raman van Wee, Chirlmin Joo*

*Corresponding author for this work

Research output: Chapter in Book/Conference proceedings/Edited volumeChapterScientificpeer-review


Fluorescence resonance energy transfer (FRET) is a photophysical phenomenon that has been repurposed as a biophysical tool to measure nanometer distances. With FRET by DNA eXchange, or FRET X, many points of interest (POIs) in a single object can be probed, overcoming a major limitation of conventional single-molecule FRET. In FRET X, short fluorescently labeled DNA imager strands specifically and transiently bind their complementary docking strands on a target molecule, such that at most a single FRET pair is formed at each point in time and multiple POIs on a single molecule can be readily probed. Here, we describe the sample preparation, image acquisition, and data analysis for structural analysis of DNA nanostructures with FRET X.

Original languageEnglish
Title of host publicationSingle Molecule Analysis
EditorsIddo Heller, David Dulin, Erwin J.G. Peterman
PublisherHumana Press Inc.
Number of pages11
ISBN (Electronic)978-1-0716-3377-9
ISBN (Print)978-1-0716-3379-3
Publication statusPublished - 2023

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Bibliographical note

Green Open Access added to TU Delft Institutional Repository 'You share, we take care!' - Taverne project
Otherwise as indicated in the copyright section: the publisher is the copyright holder of this work and the author uses the Dutch legislation to make this work public.


  • DNA nanotechnology
  • FRET X
  • Single-molecule FRET
  • Single-molecule multiplexing
  • Structural biology


Dive into the research topics of 'Single-Molecule FRET X'. Together they form a unique fingerprint.

Cite this