Apparent Concentration Quenching of Morphine Fluorescence

Brandt, Richard B.; Olsen, Marvin J.; Cheronis, Nicholas D.

doi:10.1126/science.139.3559.1063

Cited by 17 publications

(5 citation statements)

References 6 publications

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“…One well-known problem in fluorescence measurements is, however, the fluorescence inner filter effect (IFE) that can cause fluorescence spectral distortion and nonlinearity between fluorescence signal intensity and fluorophore concentration. Fluorescence IFE occurs whenever there are photon absorbers and scatterers in the sample solution, − and it is in effect even without exogenous molecular or nanoparticle-based light absorbers or scatterers. This is because fluorophores must absorb excitation photons in order to emit.…”

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confidence: 99%

“…It has been long realized that the degree of the fluorescence IFE is related to the sample UV–vis intensities at the excitation and emission wavelengths and the effective excitation and emission path lengths of the fluorophotometers. ,− ,, While UV–vis intensities of the samples can be extracted straightforwardly from the sample UV–vis spectrum, determination of the effective excitation and emission path lengths of a fluorophotometer is a nontrivial task . Figure shows a typical instrument configuration of a fluorophotometer in which the emitted photons are collected with a 90 deg angle relative to the excitation beam.…”

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Using Water Raman Intensities To Determine the Effective Excitation and Emission Path Lengths of Fluorophotometers for Correcting Fluorescence Inner Filter Effect

Nettles

Zhang

2015

Anal. Chem.

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Fluorescence and Raman inner filter effects (IFE) cause spectral distortion and nonlinearity between spectral signal intensity with increasing analyte concentration. Convenient and effective correction of fluorescence IFE has been an active research goal for decades. Presented herein is the finding that fluorescence and Raman IFE can be reliably corrected using the equation I(corr)/I(obsd) = 10(dxAx + dmAm) when the effective excitation and emission path lengths, dx and dm, of a fluorophotometer are determined by simple linear curve-fitting of Raman intensities of a series of water Raman reference samples that have known degrees of Raman IFEs. The path lengths derived with one set of Raman measurements at one specific excitation wavelength are effective for correcting fluorescence and Raman IFEs induced by any chromophore or fluorophore, regardless of the excitation and emission wavelengths. The IFE-corrected fluorescence intensities are linearly correlated to fluorophore concentration over 5 orders of magnitude (from 5.9 nM to 0.59 mM) for 2-aminopurine in a 1 cm × 0.17 cm fluorescence cuvette. This water Raman-based method is easy to implement. It does not involve complicated instrument geometry determination or difficult data manipulation. This work should be of broad significance to physical and biological sciences given the popularity of fluorescence techniques in analytical applications.

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Using Water Raman Intensities To Determine the Effective Excitation and Emission Path Lengths of Fluorophotometers for Correcting Fluorescence Inner Filter Effect

Nettles

Zhang

2015

Anal. Chem.

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“…However, it should be noted that the fluorescence intensity seemed to increase with the complex formation, suggesting that a certain fluorescence quenching in free form might affect the polarization (both the polarization intensity and the polarization potency). A previous study showed the fluorescence quenching may be influenced by factors such as solvent, van der Waals force, 26 concentration, 27 etc. It means the I ∥ − I ⊥ index is affected by not only fluorescence polarization but also fluorescence intensities, some of which may be caused by fluorescence quenching.…”

Section: Conclusion and Discussionmentioning

confidence: 99%

A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization

Zheng,

Chen,

Liu

et al. 2024

Anal. Methods

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“…Besides the collective NP surface effect, NPs also modify fluorophore fluorescence through the fluorescence inner filter effect (IFE). IFE is well-known in fluorescence and Raman spectroscopy and is attributed to the attenuation of excitation and emission light intensities induced by photon absorbers and scatterers in the sample solution. ,,,− Critically, fluorescence IFE is in play for all fluorophores in the NP-containing solution regardless of whether or not the fluorophore is adsorbed onto NPs. In addition, all NPs can impose strong IFE on fluorophore fluorescence.…”

Section: Introductionmentioning

confidence: 99%

A Generalized Model on the Effects of Nanoparticles on Fluorophore Fluorescence in Solution

Zhang

Nettles

2015

J. Phys. Chem. C

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Nanoparticles (NP) can modify fluorophore fluorescence in solution through multiple pathways that include fluorescence inner filter effect (IFE), dynamic and static quenching, surface enhancement, and fluorophore quantum yield variation associated with structural and conformational modifications induced by NP binding. The combined contribution of the latter three effects is termed the collective near-field effect because (1) they affect only fluorophore fluorescence in molecules close to the NPs, and (2) it is impossible to differentiate these effects with steady-state fluorescence measurements. A generalized model (F 0 corr /F NP corr = (1 + K[NP])/(1 + K[NP]S) was developed for the determination of the NP collective near-field effect S on the fluorophore fluorescence in the surface-adsorbed molecules. The popular Stern−Volmer equation (F 0 corr /F NP corr = (1 + K[NP]) used in current fluorescence studies of NP interfacial interactions is a special case of this generalized model, valid only under situations in which the surfacebound molecules are completely fluorescence inactive (S = 0). In addition, we excluded the possibility of NPs inducing significant dynamic fluorescence quenching under realistic experimental conditions on the basis of a simple back-of-the-envelope calculation. Furthermore, using an external reference fluorescence IFE correction method developed in this work, we demonstrated that gold nanoparticles (AuNPs) only slightly attenuate, but do not completely quench the fluorescence signal of the protein, bovine serum albumin (BSA), on AuNP. This result undermines the reliability of the BSA/AuNP binding constants calculated using the Stern−Volmer equation in earlier studies of BSA/AuNP interfacial interactions. The methodology and insights provided in this work should be of general importance for fluorescence study of nanoparticle interfacial interactions. ■ INTRODUCTIONThe effect of nanoparticles (NP) on fluorophore fluorescence can be highly complicated depending on the types of NP and the structure of fluorophore-containing molecules. Taking protein tryptophan fluorescence on plasmonic gold NPs (AuNPs) as examples, both fluorescence enhancement and quenching have been proposed in the literature, 1−9 and there are large discrepancies in the degree of fluorescence enhancement or quenching factors. One common belief is that AuNPs induce static or dynamic fluorescence quenching. 9−17 Such quenching is commonly modeled with the Stern−Volmer equation to estimate the AuNP/protein binding rate or binding affinity constants.Despite its popularity, the general applicability of the Stern− Volmer equation for fluorophore interactions with NPs is highly questionable. Indeed, the Stern−Volmer equation is applicable only in situations in which fluorophore fluorescence is completely quenched in the dynamic and static fluorophore/ quencher complex. Such a scenario is unlikely to occur for general NPs such as silica, graphene, two-dimensional nanosheets, or plasmonic AuNPs. Using protein binding to AuNPs as an ...

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Apparent Concentration Quenching of Morphine Fluorescence

Cited by 17 publications

References 6 publications

Using Water Raman Intensities To Determine the Effective Excitation and Emission Path Lengths of Fluorophotometers for Correcting Fluorescence Inner Filter Effect

Using Water Raman Intensities To Determine the Effective Excitation and Emission Path Lengths of Fluorophotometers for Correcting Fluorescence Inner Filter Effect

A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization

A Generalized Model on the Effects of Nanoparticles on Fluorophore Fluorescence in Solution

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