ArticleJournal of the American Chemical Society2007
Photobleaching pathways in single-molecule FRET experiments.
Article in Journal of the American Chemical Society, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed, 100 citations in OpenAlex.
- Benchmarking dynamic analysis methods in diffusion-based single-molecule FRET.Cell reports methods · 2026Article
- Tunable Bessel beam two-photon fluorescence microscopy for high-speed volumetric imaging of brain dynamics.eLife · 2026Article
- Three-color single-molecule fluorescence resonance energy transfer to study macromolecular dynamics.Current opinion in structural biology · 2026Review
- Polarization-engineered aberration-resilient light sheet microscopy.bioRxiv : the preprint server for biology · 2026Article
- A General Analytic Approach for Rapid Diagnostics by a Simple Algorithm for Fluorescence Single Molecule Counting.Biosensors · 2026Article
- Surface-confined FRET nanoplatform printedRSC advances · 2026Article
- Expanding Horizons in Advancements of FRET Biosensing Technologies.Biosensors · 2025Review
- Increasing the accuracy of single-molecule data analysis using tMAVEN.Biophysical journal · 2024Article
- Implementation of FRET Spectrometry Using Temporally Resolved Fluorescence: A Feasibility Study.International journal of molecular sciences · 2024Article
- Fluorescent Probes and Quenchers in Studies of Protein Folding and Protein-Lipid Interactions.Chemical record (New York, N.Y.) · 2024Review
- Increasing the accuracy of single-molecule data analysis using tMAVEN.bioRxiv : the preprint server for biology · 2024Article
- Targetable Conformationally Restricted Cyanines Enable Photon-Count-Limited Applications*.Angewandte Chemie (International ed. in English) · 2021Article
- Molecular and Spectroscopic Characterization of Green and Red Cyanine Fluorophores from the Alexa Fluor and AF Series*.Chemphyschem : a European journal of chemical physics and physical chemistry · 2021Article
- FRET-based dynamic structural biology: Challenges, perspectives and an appeal for open-science practices.eLife · 2021Review
- High-throughput smFRET analysis of freely diffusing nucleic acid molecules and associated proteins.Methods (San Diego, Calif.) · 2019Article
- Raman Spectroscopic Analysis of Signaling Molecules-Dopamine Receptors Interactions in Living Cells.ACS omega · 2018Article
- Photostable and efficient upconverting nanocrystal-based chemical sensors.Optical Materials · 2018Article
- Probing Differential Binding Mechanisms of Phenylalanine-Glycine-Rich Nucleoporins by Single-Molecule FRET.Methods in enzymology · 2018Article
- Multispot single-molecule FRET: High-throughput analysis of freely diffusing molecules.PloS one · 2017Article
- Electronic tuning of self-healing fluorophores for live-cell and single-molecule imaging.Chemical science · 2017Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
Abstract
To acquire accurate structural and dynamical information on complex biomolecular machines using single-molecule fluorescence resonance energy transfer (sm-FRET), a large flux of donor and acceptor photons is needed. To achieve such fluxes, one may use higher laser excitation intensity; however, this induces increased rates of photobleaching. Anti-oxidant additives have been extensively used for reducing acceptor's photobleaching. Here we focus on deciphering the initial step along the photobleaching pathway. Utilizing an array of recently developed single-molecule and ensemble spectroscopies and doubly labeled Acyl-CoA binding protein and double-stranded DNA as model systems, we study these photobleaching pathways, which place fundamental limitations on sm-FRET experiments. We find that: (i) acceptor photobleaching scales with FRET efficiency, (ii) acceptor photobleaching is enhanced under picosecond-pulsed (vs continuous-wave) excitation, and (iii) acceptor photobleaching scales with the intensity of only the short wavelength (donor) excitation laser. We infer from these findings that the main pathway for acceptor's photobleaching is through absorption of a short wavelength photon from the acceptor's first excited singlet state and that donor's photobleaching is usually not a concern. We conclude by suggesting the use of short pulses for donor excitation, among other possible remedies, for reducing acceptor's photobleaching in sm-FRET measurements.
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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.