ArticleBiophysical journal2016
Optimal Background Estimators in Single-Molecule FRET Microscopy.
Article in Biophysical journal, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- AID self-assembly and multistranded DNA binding drive synapsis in class switch recombination.Nucleic acids research · 2026Article
- Multiplexed single-molecule characterization at the library scale.Nature protocols · 2026Review
- DNA end configurations dictate synaptic complex formation during NHEJ-mediated end bridging.Nature communications · 2026Article
- Integration Host Factor Binds DNA Holliday Junctions.International journal of molecular sciences · 2022Article
- A dark intermediate in the fluorogenic reaction between tetrazine fluorophores andBiophysical reports · 2022Article
- A serum-stable RNA aptamer specific for SARS-CoV-2 neutralizes viral entry.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices.eLife · 2021Review
- Polymerase μ in non-homologous DNA end joining: importance of the order of arrival at a double-strand break in a purified system.Nucleic acids research · 2020Article
- The essential elements for the noncovalent association of two DNA ends during NHEJ synapsis.Nature communications · 2019Article
- Precision and accuracy of single-molecule FRET measurements-a multi-laboratory benchmark study.Nature methods · 2018Article
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3 authors.
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Abstract
Single-molecule total internal reflection fluorescence (TIRF) microscopy constitutes an umbrella of powerful tools that facilitate direct observation of the biophysical properties, population heterogeneities, and interactions of single biomolecules without the need for ensemble synchronization. Due to the low signal/noise ratio in single-molecule TIRF microscopy experiments, it is important to determine the local background intensity, especially when the fluorescence intensity of the molecule is used quantitatively. Here we compare and evaluate the performance of different aperture-based background estimators used particularly in single-molecule Förster resonance energy transfer. We introduce the general concept of multiaperture signatures and use this technique to demonstrate how the choice of background can affect the measured fluorescence signal considerably. A new, to our knowledge, and simple background estimator is proposed, called the local statistical percentile (LSP). We show that the LSP background estimator performs as well as current background estimators at low molecular densities and significantly better in regions of high molecular densities. The LSP background estimator is thus suited for single-particle TIRF microscopy of dense biological samples in which the intensity itself is an observable of the technique.
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