Several hundred-fold enhanced fluorescence from single fluorophores assembled on silver nanoparticle–dielectric–metal substrate

Ray, Krishanu; Badugu, Ramachandram; Szmacinski, Henryk; Lakowicz, Joseph R.

doi:10.1039/c5cc03581c

Cited by 19 publications

(14 citation statements)

References 23 publications

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“…Characteristics and applications of MEF have been summarized in Table 1. As reported, various kinds of metals have been used for MEF, such as gold, 21,22,34,48,49,75,100,[107][108][109][110][111][112][113] silver, 30,31,35,45,69,110,112,[114][115][116][117][118][119][120] aluminum, 121,122 copper, [123][124][125] nickel, 125,126 chromium, 125, 127 zinc 125 and so on. So far, MEF of across ultraviolet to the second near infrared wavelength range have been reported.…”

Section: Introductionmentioning

confidence: 99%

Recent progress in sensing application of metal nanoarchitecture-enhanced fluorescence

Wang

Zheng

et al. 2021

Nanoscale Adv.

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Section: Introductionmentioning

confidence: 99%

Recent progress in sensing application of metal nanoarchitecture-enhanced fluorescence

Wang

Zheng

et al. 2021

Nanoscale Adv.

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“…We used single-molecule fluorescence approaches (Michalet et al, 2006; Moerner, 2007; Ray et al, 2014, 2015; Rigler and Elson, 2001; Ulbrich and Isacoff, 2007; Xie and Dunn, 1994) to evaluate the binding stoichiometry of labeled sCD4 fusion protein (sCD4-SNAP-A488; see STAR Methods) to SOSIP.664.D7. sCD4-SNAP-A488 places a single AlexaDye-488 reporter on the C terminus of sCD4-SNAP fusion protein.…”

Section: Introductionmentioning

confidence: 99%

“…sCD4-SNAP-A488 places a single AlexaDye-488 reporter on the C terminus of sCD4-SNAP fusion protein. Compared with traditional ensemble methods, single-molecule detection (SMD) enables us to observe intermediate, pre-equilibrium states, and minor sub-populations of molecules, providing important insights into binding mechanisms and possibly revealing previously unknown interactions (Dobrowsky et al, 2008; Mason et al, 2004; Michalet et al, 2006; Ray et al, 2015; Roy et al, 2008; Xie and Dunn, 1994). Furthermore, the single-molecule photobleaching approach implemented here displays a stepwise intensity drop for individual fluorophores, and thus direct counting of reporter probes is achieved by recording the total number of steps in the intensity time trace (Tsekouras et al, 2016; Ulbrich and Isacoff, 2007).…”

Section: Introductionmentioning

confidence: 99%

Stoichiometric Analyses of Soluble CD4 to Native-like HIV-1 Envelope by Single-Molecule Fluorescence Spectroscopy

et al. 2019

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SUMMARY Analyses of HIV-1 envelope (Env) binding to CD4, and the conformational changes the interactions induce, inform the molecular mechanisms and factors governing HIV-1 infection. To address these questions, we used a single-molecule detection (SMD) approach to study the nature of reactions between soluble CD4 (sCD4) and soluble HIV-1 trimers. SMD of these reactions distinguished a mixture of one, two, or three CD4-bound trimer species. Single-ligand trimers were favored at early reaction times and ligand-saturated trimers later. Furthermore, some trimers occupied by one sCD4 molecule did not bind additional ligands, whereas the majority of two ligand-bound species rapidly transitioned to the saturated state. Quantification of liganded trimers observed in reactions with various sCD4 concentrations reflected an overall negative cooperativity in ligand binding. Collectively, our results highlight the general utility of SMD in studying protein interactions and provide critical insights on the nature of sCD4-HIV-1 Env interactions.

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“…For physical adsorption isotherm studies, glass slides with Ag and different thickness of C 60 were incubated in different concentrations of RhB in de-ionized water (0.1 M, 0.01 M, 1 mM, 0.1 mM, 0.01 mM) for 24 hours after which they were taken out and washed in de-ionized water thrice and dried in an air stream. For studies with b-BSA, substrates were prepared as detailed in the study by Ray et al 27 Briefly, ~2.5 µM b-BSA solution was made in 10 mM PBS (Sigma-Aldrich) solution. Glass, glass-Ag, C 60 , and glass-Ag-C 60 substrates were then incubated in this solution for 24 hours at 51°C.…”

Section: Methodsmentioning

confidence: 99%

Chemiplasmonics for high-throughput biosensors

Raghavendra

Zhu

Gregory

et al. 2018

IJN

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Background: The sensitivity of ELISA for biomarker detection can be significantly increased by integrating fluorescence with plasmonics. In surface-plasmon-coupled emission, the fluorophore emission is generally enhanced through the so-called physical mechanism due to an increase in the local electric field. Despite its fairly high enhancement factors, the use of surface-plasmoncoupled emission for high-throughput and point-of-care applications is still hampered due to the need for expensive focusing optics and spectrometers. Methods: Here, we describe a new chemiplasmonic-sensing paradigm for enhanced emission through the molecular interactions between aromatic dyes and C 60 films on Ag substrates. Results: A 20-fold enhancement in the emission from rhodamine B-labeled biomolecules can be readily elicited without quenching its red color emission. As a proof of concept, we demonstrate two model bioassays using: 1) the RhB-streptavidin and biotin complexes in which the dye was excited using an inexpensive laser pointer and the ensuing enhanced emission was recorded by a smartphone camera without the need for focusing optics and 2) high-throughput 96-well plate assay for a model antigen (rabbit immunoglobulin) that showed detection sensitivity as low as 6.6 pM. Conclusion: Our results show clear evidence that chemiplasmonic sensors can be extended to detect biomarkers in a point-of-care setting through a smartphone in simple normal incidence geometry without the need for focusing optics. Furthermore, chemiplasmonic sensors also facilitate high-throughput screening of biomarkers in the conventional 96-well plate format with 10-20 times higher sensitivity.

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Several hundred-fold enhanced fluorescence from single fluorophores assembled on silver nanoparticle–dielectric–metal substrate

Cited by 19 publications

References 23 publications

Recent progress in sensing application of metal nanoarchitecture-enhanced fluorescence

Recent progress in sensing application of metal nanoarchitecture-enhanced fluorescence

Stoichiometric Analyses of Soluble CD4 to Native-like HIV-1 Envelope by Single-Molecule Fluorescence Spectroscopy

Chemiplasmonics for high-throughput biosensors

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