ArticleProceedings of SPIE--the International Society for Optical Engineering2013
8-spot smFRET analysis using two 8-pixel SPAD arrays.
Article in Proceedings of SPIE--the International Society for Optical Engineering, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- An automated single-molecule FRET platform for high-content, multiwell plate screening of biomolecular conformations and dynamics.Nature communications · 2023Article
- Fast Gating for Raman Spectroscopy.Sensors (Basel, Switzerland) · 2021Review
- High-throughput smFRET analysis of freely diffusing nucleic acid molecules and associated proteins.Methods (San Diego, Calif.) · 2019Article
- 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
- 48-spot single-molecule FRET setup with periodic acceptor excitation.The Journal of chemical physics · 2018Article
- AnThe Journal of biological chemistry · 2017Article
- 16-Ch Time-resolved Single-Molecule Spectroscopy Using Line Excitation.Proceedings of SPIE--the International Society for Optical Engineering · 2017Article
- Multispot single-molecule FRET: High-throughput analysis of freely diffusing molecules.PloS one · 2017Article
- Photon-HDF5: Open Data Format and Computational Tools for Timestamp-based Single-Molecule Experiments.Proceedings of SPIE--the International Society for Optical Engineering · 2016Article
- FRETBursts: An Open Source Toolkit for Analysis of Freely-Diffusing Single-Molecule FRET.PloS one · 2016Article
- Silicon photon-counting avalanche diodes for single-molecule fluorescence spectroscopy.IEEE journal of selected topics in quantum electronics : a publication of the IEEE Lasers and Electro-optics Society · 2014Article
- smFRET studies of the 'encounter' complexes and subsequent intermediate states that regulate the selectivity of ligand binding.FEBS letters · 2014Review
- A 48-pixel array of Single Photon Avalanche Diodes for multispot Single Molecule analysis.Proceedings of SPIE--the International Society for Optical Engineering · 2013Article
- Single-molecule FRET experiments with a red-enhanced custom technology SPAD.Proceedings of SPIE--the International Society for Optical Engineering · 2013Article
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Authors and funding
8 authors.
Funding
Abstract
Single-molecule Förster resonance energy transfer (smFRET) techniques are now widely used to address outstanding problems in biology and biophysics. In order to study freely diffusing molecules, current approaches consist in exciting a low concentration (<100 pM) sample with a single confocal spot using one or more lasers and detecting the induced single-molecule fluorescence in one or more spectrally- and/or polarization-distinct channels using single-pixel Single-Photon Avalanche Diodes (SPADs). A large enough number of single-molecule bursts must be accumulated in order to compute FRET efficiencies with sufficient statistics. As a result, the minimum timescale of observable phenomena is set by the minimum acquisition time needed for accurate measurements, typically a few minutes or more, limiting this approach mostly to equilibrium studies. Increasing smFRET analysis throughput would allow studying dynamics with shorter timescales. We recently demonstrated a new multi-spot excitation approach, employing a novel multi-pixel SPAD array, using a simplified dual-view setup in which a single 8-pixel SPAD array was used to collect FRET data from 4 independent spots. In this work we extend our results to 8 spots and use two 8-SPAD arrays to collect donor and acceptor photons and demonstrate the capabilities of this system by studying a series of doubly labeled dsDNA samples with different donor-acceptor distances ranging from low to high FRET efficiencies. Our results show that it is possible to enhance the throughput of smFRET measurements in solution by almost one order of magnitude, opening the way for studies of single-molecule dynamics with fast timescale once larger SPAD arrays become available.
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