Ultraviolet Photostability Improvement for Autofluorescence Correlation Spectroscopy on Label-Free Proteins

Barulin, Aleksandr; Wenger, Jérôme

doi:10.1021/acs.jpclett.0c00209

Cited by 16 publications

(33 citation statements)

References 73 publications

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“…Tryptophan emission in proteins is often inherently quenched due to electron transfers to the neighboring amino acids or energy homotransfer among the tryptophan residues in the protein structure [18,35]. Thus, β-galactosidase and streptavidin possess low quantum yields of 1.6% and 3.5%, respectively [36].…”

Section: Uv Dye and Protein Samplesmentioning

confidence: 99%

Purcell radiative rate enhancement of label-free proteins with ultraviolet aluminum plasmonics

Barulin

Roy

Claude

et al. 2021

J. Phys. D: Appl. Phys.

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The vast majority of proteins are intrinsically fluorescent in the ultraviolet, thanks to the emission from their tryptophan and tyrosine amino-acid constituents. However, the protein autofluorescence quantum yields are generally very low due to the prevailing quenching mechanisms by other amino acids inside the protein. This motivates the interest to enhance the radiative emission rate of proteins using nanophotonic structures. Although there have been numerous reports of Purcell effect and local density of optical states control in the visible range using single dipole quantum emitters, the question remains open to apply these concepts in the UV on real proteins containing several tryptophan and tyrosine amino acids arranged in a highly complex manner. Here, we report the first complete characterization of the Purcell effect and radiative rate enhancement for the UV intrinsic fluorescence of label-free β-galactosidase and streptavidin proteins in plasmonic aluminum nanoapertures. We find an excellent agreement with a calibration performed using a high quantum yield UV fluorescent dye. Demonstrating and intensifying the Purcell effect is essential for the applications of UV plasmonics and the label-free detection of single proteins.

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Section: Uv Dye and Protein Samplesmentioning

confidence: 99%

Purcell radiative rate enhancement of label-free proteins with ultraviolet aluminum plasmonics

Barulin

Roy

Claude

et al. 2021

J. Phys. D: Appl. Phys.

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“…(c) Super-resolved in cellulo biochemistry (adapted from [83]). (d) Enhancing label-free UV fluorescence of proteins with ZWGs (adapted from [44]). (e) Artist view of a SPAD array (adapted from [196]).…”

Section: Discussionmentioning

confidence: 99%

“…The combination of expertise from biophysics and nanophotonics turned out to be particularly fruitful for addressing this challenge. Indeed, detecting the very weak signal coming from protein UV autofluorescence was recently demonstrated thanks to the use of aluminum ZWGs [44] (see Figure 12d). The interaction of a single protein with the ZWG enhances protein autofluorescence and enables detection of proteins at physiological concentrations.…”

Section: Discussionmentioning

confidence: 99%

“…Interestingly, the use of subwavelength ZWGs can strongly influence the FRET efficiency [39][40][41], allowing for the observation of FRET at distances that were previously inaccessible [42] and for relative orientations between molecules for which FRET would otherwise be forbidden [43]. Furthermore, it has very recently been shown that the strong fluorescence enhancement generated by rectangle-shaped aluminum ZWGs makes possible the observation of the autofluorescence of single unlabeled proteins emitting very weakly in the UV region of the spectrum [44]. This opens very interesting perspectives in biophysics, allowing the optical detection of proteins without the requirement of potentially disturbing external fluorescent labeling.…”

Section: Introductionmentioning

confidence: 99%

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Super-resolution imaging: when biophysics meets nanophotonics

et al. 2021

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Probing light–matter interaction at the nanometer scale is one of the most fascinating topics of modern optics. Its importance is underlined by the large span of fields in which such accurate knowledge of light–matter interaction is needed, namely nanophotonics, quantum electrodynamics, atomic physics, biosensing, quantum computing and many more. Increasing innovations in the field of microscopy in the last decade have pushed the ability of observing such phenomena across multiple length scales, from micrometers to nanometers. In bioimaging, the advent of super-resolution single-molecule localization microscopy (SMLM) has opened a completely new perspective for the study and understanding of molecular mechanisms, with unprecedented resolution, which take place inside the cell. Since then, the field of SMLM has been continuously improving, shifting from an initial drive for pushing technological limitations to the acquisition of new knowledge. Interestingly, such developments have become also of great interest for the study of light–matter interaction in nanostructured materials, either dielectric, metallic, or hybrid metallic-dielectric. The purpose of this review is to summarize the recent advances in the field of nanophotonics that have leveraged SMLM, and conversely to show how some concepts commonly used in nanophotonics can benefit the development of new microscopy techniques for biophysics. To this aim, we will first introduce the basic concepts of SMLM and the observables that can be measured. Then, we will link them with their corresponding physical quantities of interest in biophysics and nanophotonics and we will describe state-of-the-art experiments that apply SMLM to nanophotonics. The problem of localization artifacts due to the interaction of the fluorescent emitter with a resonant medium and possible solutions will be also discussed. Then, we will show how the interaction of fluorescent emitters with plasmonic structures can be successfully employed in biology for cell profiling and membrane organization studies. We present an outlook on emerging research directions enabled by the synergy of localization microscopy and nanophotonics.

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“…In this article, we did not discuss the role of oxide on performance, as quite a lot of work is going on to solve this issue. [28,29] The general idea presented in the article is the use of a combination of feeder and reflector antenna to overcome drawbacks when they are used in a standalone situation. Analogous to our modeled structure in the microwave regime is the dish antenna, which uses horn antennas as a feeder (gap antenna in our case) and parabolic antenna as a reflector.…”

Section: Discussionmentioning

confidence: 99%

A Highly Directive Ultraviolet Plasmonic “Antenna‐on‐Reflector” for Single‐Molecule Detection

Roy

Bolshakov

2021

Physica Rapid Research Ltrs

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Single‐molecule detection (SMD) has been a hot topic for decades due to its extensive implementation in biomedical, chemical, physical, and material sciences. The methods used for single‐molecule detection are generally variants of fluorescence and Raman spectroscopy, which employs plasmonic particles for hot‐spot generation upon illumination. The signal generated by molecules in the hot spot is strong but not directive, so, <in most contemporary works, a nanomolar concentration is required to suppress noise from background molecules. However, at extremely low concentrations, biological samples vary with regard to conformation and interaction dynamics, so the most suitable dilution is micromolar. Thus an optical antenna that can both focus incident light at tight spots and be highly directive for the efficient collection of fluorescence is demanded. Herein, a nanoantenna in the shape of an aluminium hemisphere on a parabolic reflector providing both incident wave enhancement and efficient directive signal outcoupling is proposed and simulated. To achieve this feat, the focus of the parabolic reflector coincides spatially with the hot spot created by the plasmonic gap antenna. The structure shows broadband () escalation in directivity, 24–28 dB. The calculated incident field enhancement in the detection volume of the plasmonic gap is up to .

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Ultraviolet Photostability Improvement for Autofluorescence Correlation Spectroscopy on Label-Free Proteins

Cited by 16 publications

References 73 publications

Purcell radiative rate enhancement of label-free proteins with ultraviolet aluminum plasmonics

Purcell radiative rate enhancement of label-free proteins with ultraviolet aluminum plasmonics

Super-resolution imaging: when biophysics meets nanophotonics

A Highly Directive Ultraviolet Plasmonic “Antenna‐on‐Reflector” for Single‐Molecule Detection

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