ArticlePloS one2017
Multispot single-molecule FRET: High-throughput analysis of freely diffusing molecules.
Article in PloS one, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Who cites it
17 citing papers in PubMed.
- Single-molecule detection and super-resolution imaging with a portable and adaptable 3D-printed microscopy platform (Brick-MIC).Science advances · 2024Article
- Single-molecule detection and super-resolution imaging with a portable and adaptable 3D-printed microscopy platform (Brick-MIC).bioRxiv : the preprint server for biology · 2024Article
- Multi-parameter photon-by-photon hidden Markov modeling.Nature communications · 2022Article
- The structural heterogeneity of α-synuclein is governed by several distinct subpopulations with interconversion times slower than milliseconds.Structure (London, England : 1993) · 2021Article
- Utilizing Time-Resolved Protein-Induced Fluorescence Enhancement to Identify Stable Local Conformations One α-Synuclein Monomer at a Time.Journal of visualized experiments : JoVE · 2021Article
- FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices.eLife · 2021Review
- Custom silicon technology for SPAD-arrays with red-enhanced sensitivity and low timing jitter.Optics express · 2021Article
- High-throughput smFRET analysis of freely diffusing nucleic acid molecules and associated proteins.Methods (San Diego, Calif.) · 2019Article
- Systematic Assessment of Burst Impurity in Confocal-Based Single-Molecule Fluorescence Detection Using Brownian Motion Simulations.Molecules (Basel, Switzerland) · 2019Article
- Article
- Single-photon avalanche diode imagers in biophotonics: review and outlook.Light, science & applications · 2019Review
- Optical crosstalk in SPAD arrays for high-throughput single-molecule fluorescence spectroscopy.Nuclear instruments & methods in physics research. Section A, Accelerators, spectrometers, detectors and associated equipment · 2018Article
- Characterizing highly dynamic conformational states: The transcription bubble in RNAP-promoter open complex as an example.The Journal of chemical physics · 2018Article
- 48-spot single-molecule FRET setup with periodic acceptor excitation.The Journal of chemical physics · 2018Article
- Toward dynamic structural biology: Two decades of single-molecule Förster resonance energy transfer.Science (New York, N.Y.) · 2018Review
- Different types of pausing modes during transcription initiation.Transcription · 2017Article
- A user-friendly tool to convert photon counting data to the open-source Photon-HDF5 file format.Proceedings of SPIE--the International Society for Optical EngineeringArticle
Corrections and comments
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Authors and funding
9 authors.
Funding
Abstract
We describe an 8-spot confocal setup for high-throughput smFRET assays and illustrate its performance with two characteristic experiments. First, measurements on a series of freely diffusing doubly-labeled dsDNA samples allow us to demonstrate that data acquired in multiple spots in parallel can be properly corrected and result in measured sample characteristics consistent with those obtained with a standard single-spot setup. We then take advantage of the higher throughput provided by parallel acquisition to address an outstanding question about the kinetics of the initial steps of bacterial RNA transcription. Our real-time kinetic analysis of promoter escape by bacterial RNA polymerase confirms results obtained by a more indirect route, shedding additional light on the initial steps of transcription. Finally, we discuss the advantages of our multispot setup, while pointing potential limitations of the current single laser excitation design, as well as analysis challenges and their solutions.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.