2023
DOI: 10.1002/adom.202300168
|View full text |Cite
|
Sign up to set email alerts
|

Achieving High Temporal Resolution in Single‐Molecule Fluorescence Techniques Using Plasmonic Nanoantennas

Abstract: Single‐molecule fluorescence techniques are essential for investigating the molecular mechanisms in biological processes. However, achieving sub‐millisecond temporal resolution to monitor fast molecular dynamics remains a significant challenge. The fluorescence brightness is the key parameter that generally defines the temporal resolution for these techniques. Conventional microscopes and standard fluorescent emitters fall short in achieving the high brightness required for sub‐millisecond monitoring. Plasmoni… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
4

Citation Types

0
4
0

Year Published

2023
2023
2024
2024

Publication Types

Select...
4
1

Relationship

0
5

Authors

Journals

citations
Cited by 5 publications
(4 citation statements)
references
References 106 publications
(177 reference statements)
0
4
0
Order By: Relevance
“…Crucially, although methods enabling access to faster time scales through correlation techniques exist, , time-resolving individual rare events, such as the rapid jumps across the barriers between two conformational states (e.g., protein or nucleic acid folding), require very high count rates from single molecules. , These transition paths have gained increasing interest, and smFRET experiments have been playing a major role in revealing the nature and time scales of these paths. , However, the photon count rate (PCR) required for accessing the relevant microsecond time scale mandates the use of high excitation intensities, leading to saturation and increased photobleaching, which makes the task of measuring these transition paths very challenging. Plasmonic hotspots have been shown to increase the photostability and brightness of fluorescent labels , and offer a potentially complementary strategy to the more commonly used chemical photostabilization. Coupling the emitters to plasmonic hotspots not only increases the electric field between the two plasmonic nanoparticles but also increases the emission rates of the fluorophore, leading to a shorter time that the molecule spends in the reactive excited states as well as reducing the time until the fluorophore is available for re-excitation. , This, in turn, results in improved photostability of fluorescent labels and also enables higher fluorescence intensities without saturation. ,, First examples employing similar strategies for diffusing molecules have appeared and showed promising results, , but they are limited to the submillisecond time scales of molecular diffusion through the excitation volume, which makes the complete observation of complex biomolecular pathways unlikely. DNA origami nanoantennas , have overcome the challenge of selective immobilization of biomolecules in plasmonic hotspots.…”
Section: Introductionmentioning
confidence: 99%
See 1 more Smart Citation
“…Crucially, although methods enabling access to faster time scales through correlation techniques exist, , time-resolving individual rare events, such as the rapid jumps across the barriers between two conformational states (e.g., protein or nucleic acid folding), require very high count rates from single molecules. , These transition paths have gained increasing interest, and smFRET experiments have been playing a major role in revealing the nature and time scales of these paths. , However, the photon count rate (PCR) required for accessing the relevant microsecond time scale mandates the use of high excitation intensities, leading to saturation and increased photobleaching, which makes the task of measuring these transition paths very challenging. Plasmonic hotspots have been shown to increase the photostability and brightness of fluorescent labels , and offer a potentially complementary strategy to the more commonly used chemical photostabilization. Coupling the emitters to plasmonic hotspots not only increases the electric field between the two plasmonic nanoparticles but also increases the emission rates of the fluorophore, leading to a shorter time that the molecule spends in the reactive excited states as well as reducing the time until the fluorophore is available for re-excitation. , This, in turn, results in improved photostability of fluorescent labels and also enables higher fluorescence intensities without saturation. ,, First examples employing similar strategies for diffusing molecules have appeared and showed promising results, , but they are limited to the submillisecond time scales of molecular diffusion through the excitation volume, which makes the complete observation of complex biomolecular pathways unlikely. DNA origami nanoantennas , have overcome the challenge of selective immobilization of biomolecules in plasmonic hotspots.…”
Section: Introductionmentioning
confidence: 99%
“…14,15 This, in turn, results in improved photostability of fluorescent labels and also enables higher fluorescence intensities without saturation. 9,16,17 First examples employing similar strategies for diffusing molecules have appeared and showed promising results, 18,19 but they are limited to the submillisecond time scales of molecular diffusion through the excitation volume, which makes the complete observation of complex biomolecular pathways unlikely. DNA origami nanoantennas 20,21 have overcome the challenge of selective immobilization of biomolecules in plasmonic hotspots.…”
Section: ■ Introductionmentioning
confidence: 99%
“…Plasmonic nanoparticles are revolutionizing the field because they can enhance a dye’s fluorescence intensity >10 3 -fold resulting in a vastly improved signal-to-background ratio. These have been employed to probe single biomolecules in highly concentrated samples , and to improve the overall brightness of single fluorophores. Plasmon-enhanced fluorescence has recently been applied to the field of diagnostics, , where multiple assay designs were used to detect analyte in point-of-care devices. The strong plasmon-enhanced signals generated by a dimeric nanogap antenna enabled miniaturization of the optical setup to a point-of-care device, thereby highlighting the promise of the technology for biosensing.…”
Section: Introductionmentioning
confidence: 99%
“…This, in turn, results in improved photostability of fluorescent labels and also enables higher fluorescence intensities without saturation [9, 16, 17]. First examples employing similar strategies for diffusing molecules have appeared and showed promising results [18, 19], but they are limited to the submillisecond timescales of molecular diffusion through the excitation volume, which makes the complete observation of complex biomolecular pathways unlikely. DNA origami nanoantennas [20, 21] have overcome the challenge of selective immobilization of biomolecules in plasmonic hotspots.…”
Section: Introductionmentioning
confidence: 99%